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FGF23 is a cardiovascular toxin in CKD

Chronic kidney disease (CKD) is a serious health problem that involves gradual loss of kidney function. The major cause of death in CKD patients is cardiovascular disease is triggered by the cardiovascular toxins fibroblast growth factor (FGF23) and phosphate.

Declining kidney  function leads to elevated  serum phosphate levels which are regulated by the phophaturic hormone FGF23 in CKD patients.  In later stages of chronic kidney disease, access FGF23 and phosphate levels lead to increased cardiovascular disease and mortality.  Ongoing experimental studies are identifying  novel therapeutic strategies in order to prevent toxicity of FGF23 and hyperphosphatemia in CKD patients.

FGF23 is a cardiovascular toxin in chronic kidney disease

FGF23 and Phosphate-Cardiovascular Toxins in CKD.

Vogt I, Haffner D, Leifheit-Nestler M. Toxins (Basel). 2019 Nov 6;11(11):647. doi: 10.3390/toxins11110647. PMID: 31698866; PMCID: PMC6891626.

 Biomedica offers ready to use FGF23 (C-terminal) and FGF23 intact ELISA kits                     

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FGF23 and Phosphate-Cardiovascular Toxins in CKD  full text link

Abstract

Elevated levels of fibroblast growth factor 23 (FGF23) and phosphate are highly associated with increased cardiovascular disease and mortality in patients suffering from chronic kidney disease (CKD). As the kidney function declines, serum phosphate levels rise and subsequently induce the secretion of the phosphaturic hormone FGF23. In early stages of CKD, FGF23 prevents the increase of serum phosphate levels and thereby attenuates phosphate-induced vascular calcification, whereas in end-stage kidney disease, FGF23 fails to maintain phosphate homeostasis. Both hyperphosphatemia and elevated FGF23 levels promote the development of hypertension, vascular calcification, and left ventricular hypertrophy by distinct mechanisms. Therefore, FGF23 and phosphate are considered promising therapeutic targets to improve the cardiovascular outcome in CKD patients. Previous therapeutic strategies are based on dietary and pharmacological reduction of serum phosphate, and consequently FGF23 levels. However, clinical trials proving the effects on the cardiovascular outcome are lacking. Recent publications provide evidence for new promising therapeutic interventions, such as magnesium supplementation and direct targeting of phosphate and FGF receptors to prevent toxicity of FGF23 and hyperphosphatemia in CKD patients.

RELATED PUBLICATIONS

 Fibroblast Growth Factor-23-A Potential Uremic Toxin.

Kuczera P, Adamczak M, Wiecek A Toxins (Basel). 2016 Dec 8;8(12):369. doi: 10.3390/toxins8120369. PMID: 27941640; PMCID: PMC5198563.

 Abstract

Fibroblast growth factor-23 (FGF23) is a circulating member of the FGF family produced mainly by the osteocytes and osteoblasts that can act as a hormone. The main action of FGF23 is to lower phosphatemia via the reduction of urinary phosphate reabsorption and the decrease of 1,25(OH)₂-D generation in the kidney. In the course of chronic kidney disease (CKD), plasma FGF23 concentration rises early, most probably to compensate the inability of the deteriorating kidneys to excrete an adequate amount of phosphate. However, this comes at the cost of FGF23-related target organ toxicity. Results of clinical studies suggest that elevated plasma FGF23 concentration is independently associated with the increased risk of CKD progression, occurrence of cardio-vascular complications, and mortality in different stages of CKD. FGF23 also contributes to cardiomyocyte hypertrophy, vascular calcification, and endothelial dysfunction. The impact of FGF23 on heart muscle is not dependent on Klotho, but rather on the PLCγ-calcineurin-NFAT (nuclear factor of activated T-cells) pathway. Among the factors increasing plasma FGF23 concentration, active vitamin D analogues play a significant role. Additionally, inflammation and iron deficiency can contribute to the increase of plasma FGF23. Among the factors decreasing plasma FGF23, dietary phosphate restriction, some intestinal phosphate binders, cinacalcet (and other calcimimetics), and nicotinamide can be enumerated. Anti-FGF23 antibodies have also recently been developed to inhibit the action of FGF23 in target organs. Still, the best way to normalize plasma FGF23 in maintenance hemodialysis patients is restoring kidney function by successful kidney transplantation.

Effect of different phosphate binders on fibroblast growth factor 23 levels in patients with chronic kidney disease: a systematic review and meta-analysis of randomized controlled trials.

Zhao SJ, Wang ZX, Chen L, Wang FX, Kong LD. Ann Palliat Med. 2022 Apr;11(4):1264-1277. doi: 10.21037/apm-21-1943. Epub 2021 Nov 2. PMID: 34775773.

 Simultaneous management of disordered phosphate and iron homeostasis to correct fibroblast growth factor 23 and associated outcomes in chronic kidney disease.

Courbon G, Martinez-Calle M, David V. Curr Opin Nephrol Hypertens. 2020 Jul;29(4):359-366. doi: 10.1097/MNH.0000000000000614. PMID: 32452919; PMCID: PMC7769207.

Leucine-rich alpha-2 glycoprotein – an inflammation marker

LRG – a biomarker of fetal infection: Infections during pregnancy could harm the pregnant mother and the developing baby and remain a substantial clinical problem. Findings suggest that certain biomarkers have diagnostic value when maternal infection is suspected. Researchers from Japan recently identified the inflammatory protein leucine-rich alpha-2 glycoprotein (LRG) as a biomarker of fetal infection. Read more: Evaluation of leucine-rich alpha-2 glycoprotein as a biomarker of fetal infection.

LRG can reliably be measured by ELISA assay in serum, plasma, urine or cell culture supernatants.

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Check out the LRG ELISA assay validation data

RELATED PUBLICATIONS

Evaluation of leucine-rich alpha-2 glycoprotein as a biomarker of fetal infection.  

Kajimoto E, Endo M, Fujimoto M, Matsuzaki S, Fujii M, Yagi K, Kakigano A, Mimura K, Tomimatsu T, Serada S, Takeuchi M, Yoshino K, Ueda Y, Kimura T, Naka T.

PLoS One. 2020, 19;15(11):e0242076. doi: 10.1371/journal.pone.0242076. PMID: 33211747; PMCID: PMC7676652. PubMed

Abstract

This study aimed to determine the association between umbilical cord leucine-rich alpha-2 glycoprotein (LRG) and fetal infection and investigate the underlying mechanism of LRG elevation in fetuses. We retrospectively reviewed the medical records of patients who delivered at Osaka University Hospital between 2012 and 2017 and selected those with histologically confirmed chorioamnionitis (CAM), which is a common pregnancy complication that may cause neonatal infection. The participants were divided into two groups: CAM with fetal infection (CAM-f[+] group, n = 14) and CAM without fetal infection (CAM-f[-] group, n = 31). Fetal infection was defined by the histological evidence of funisitis. We also selected 50 cases without clinical signs of CAM to serve as the control. LRG concentrations in sera obtained from the umbilical cord were unaffected by gestational age at delivery, neonatal birth weight, nor the presence of noninfectious obstetric complications (all, p > 0.05). Meanwhile, the LRG levels (median, Interquartile range [IQR]) were significantly higher in the CAM-f(+) group (10.37 [5.21-13.7] μg/ml) than in the CAM-f(-) (3.61 [2.71-4.65] μg/ml) or control group (3.39 [2.81-3.93] μg/ml; p < 0.01). The area under the receiver operating characteristic (ROC) curve of LRG for recognizing fetal infection was 0.92 (optimal cutoff, 5.08 μg/ml; sensitivity, 86%; specificity, 88%). In a mouse CAM model established by lipopolysaccharide administration, the fetal LRG protein in sera and LRG mRNA in the liver were significantly higher than those in phosphate-buffered saline (PBS)-administered control mice (p < 0.01). In vitro experiments using a fetal liver-derived cell line (WRL68) showed that the expression of LRG mRNA was significantly increased after interleukin (IL)-6, IL-1β, and tumor necrosis factor- alpha (TNF-α) stimulation (p < 0.01); the induction was considerably stronger following IL-6 and TNF-α stimulation (p < 0.01). In conclusion, LRG is an effective biomarker of fetal infection, and fetal hepatocytes stimulated with inflammatory cytokines may be the primary source of LRG production in utero.

Recognising the burden of maternal infection worldwide.

Seale AC.  Lancet Glob Health. 2020. 8(5):e615-e616. doi: 10.1016/S2214-109X(20)30126-1. PMID: 32353299.

 A systematic review of biomarkers associated with maternal infection in pregnant and postpartum women.

Oben AG, Johnson BM, Tita ATN, Andrews WW, Hibberd PL, Subramaniam A, Sinkey RG. Int J Gynaecol Obstet. 2022.157(1):42-50. doi: 10.1002/ijgo.13747. PMID: 33999419.

 An update on COVID-19 and pregnancy.

Jamieson DJ, Rasmussen SA. Am J Obstet Gynecol. 2022. 226(2):177-186. doi: 10.1016/j.ajog.2021.08.054. PMID: 34534497; PMCID: PMC8438995.

Developed & manufactured by Biomedica – Biomarkers for Cancer Research

The identification and validation of biomarkers in cancer is essential to improve our understanding of the disease. The emergence of novel cancer biomarkers continues to grow as scientists strive to find promising novel therapeutic targets and new prognostic and predictive markers to fight the disease.

Discover Biomedica Biomarker Assays for Cancer Research – we offer a range of unique biomarker ELISAs for your research.

Advantages of Biomedica ELISA kits

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• LOW sample volumes
• TRUSTED – widely cited

High specificity – known target binding sites through mapping data
The unique specificity of the proprietary antibodies used in the Biomedica ELISA kits ensure that the assays only measure the analyte of interest.

Discover Biomedica´s Biomarker ELISA kits for cancer research
Neuropilin-1, Periostin, Semaphorin 4D, Osteoprotegerin, RANKL, LRG1, IL-6, VEGF, Angiopoietin-2 and other

Biomarkers for Cancer Research – Learn more about the markers

The transmembrane protein Neuropilin-1 (NRP1) regulates tumor biology and has been identified as a checkpoint target (1). High tissue NRP-1 levels are associated with a poor prognosis in breast cancer patients. In a recent study (2), German researchers have shown that circulating soluble NRP1 serum levels are an independent marker for poor prognosis in early breast cancer. Soluble Neuropilin-1 was quantified in serum with the highly specific NRP1 ELISA from Biomedica. Therapeutic areas of NRP1.

The secreted extracellular matrix protein Periostin has evolved as a novel therapeutic target and is a robust marker of glioma malignancy and potential tumor recurrence. It has also been implicated in the pathogenesis of breast cancer as high serum levels of periostin are associated with a poor survival in breast cancer patients (3). Periostin was quantified in serum with the well characterized Biomedica Periostin ELISA that has been published (4). Therapeutic areas of Periostin.

Semaphorin 4D (Sema4D) is a glycoprotein that is emerging as clinical biomarker and as therapeutic target in cancer. It has been associated with cancer progression and the occurrence of bone metastases (5, 6). Therapeutic areas of Sema4D.

Leucine-rich alpha-2-glycoprotein 1 (LRG1) is a protein that is an important factor involved in pathogenic angiogenesis in cancer. It is abundantly present in the microenvironment of many tumors contributing to vascular dysfunction and thus serving as a potential therapeutic target (7). Therapeutic areas of LRG 1.

The RANKL/RANK/OPG system contributes to the development of bone metastases and influences tumor biology in earlier stages of cancer (9). Dysregulation has been widely documented in the context of metastatic bone disease (10). The Biomedica OPG and RANKL ELISA kits have been widely used in the respective studies.

Literature
1. Neuropilin-1: a checkpoint target with unique implications for cancer immunology and immunotherapy. Chuckran CA et al., J Immunother Cancer. 2020. 8(2):e000967.
2. Soluble Neuropilin-1 is an independent marker of poor prognosis in early breast cancer. Rachner TD et al., J Cancer Res Clin Oncol. 2021. 147(8):2233-2238.
3. High serum levels of periostin are associated with a poor survival in breast cancer. Rachner TD et al., 2020. 180(2):515-524.
4. Characterization of a sandwich ELISA for the quantification of all human periostin isoforms. Gadermaier E J Clin Lab Anal. 2018. 32(2):e22252.
5. Plasma levels of Semaphorin 4D are decreased by adjuvant tamoxifen but not aromatase inhibitor therapy in breast cancer patients. Göbel A J Bone Oncol. 2019. 4;16:100237.
6. Semaphorins as emerging clinical biomarkers and therapeutic targets in cancer. Mastrantonio R et al., Theranostics. 202. 15;11(7):3262-3277.
7. Leucine-rich alpha-2-glycoprotein 1 (LRG1) as a novel ADC target. Javaid F et al., RSC Chem Biol. 2021. 31;2(4):1206-1220.
8. RANKL/RANK/OPG system beyond bone remodeling: involvement in breast cancer and clinical perspectives. Infante M J Exp Clin Cancer Res. 2019. 8;38(1):12.
9. Serum receptor activator of nuclear factor κB ligand (RANKL) levels predict biochemical recurrence in patients undergoing radical prostatectomy. Todenhöfer T, BJU Int. 2014. 113(1):152-9.
10. Prognostic Value of RANKL/OPG Serum Levels and Disseminated Tumor Cells in Nonmetastatic Breast Cancer. Rachner TD et al., Clin Cancer Res. 2019. 15;25(4):1369-1378.

 

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We validate all the BIOMEDICA ELISA assays  according to international quality standards and follow guidelines of ICH, EMEA and FDA. Thus, you can find the validation data of every assay on the respective product pages on our website. Specifically, the data include spike-recovery and parallelism, dilution linearity experiments including precision and accuracy testing. In addition, every analyte is tested on its stability in the respective sample matrix, including long term stability. Finally, the antibodies utilized in the BIOMEDICA Elisa kits are characterized and epitope mapped. Click here to upload the validation data file on our human Interleukin-6 (IL-6) ELISA assay kit. 

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Related Posts – on Biomedica Quality Cytokine Kits

All our ELISA kits undergo a stringent quality control process, ensuring reliable and consistent results.  We are located right in the heart of Europe.  developed & manufactured by Biomedica.

 

 

Report on novel prostate tumor cell targets

 The Biomedica EZ4U – Cell Proliferation & Cytotoxicity Kit was cited in a high tier journal studying novel prostate tumor endothelial cell targets. The study highlights  CXCR4/CXCL12 interaction as a potential novel target to interfere with tumor angiogenesis:

Comprehensive characterization of the prostate tumor microenvironment identifies CXCR4/CXCL12 crosstalk as a novel antiangiogenic therapeutic target in prostate cancer.

EZ4U Cell Proliferation & Cytotoxicity Assay

EZ4U – Test Highlights

• Use of non-radioactive & non-toxic substances
• Easy one-step procedure on living cells
• Cited in more than 225 publications

More information can be found in our brochure:  EZ4U – cat. no. BI-5000

Comprehensive characterization of the prostate tumor microenvironment identifies CXCR4/CXCL12 crosstalk as a novel antiangiogenic therapeutic target in prostate cancer.

Heidegger I, Fotakis G, Offermann A, Goveia J, Daum S, Salcher S, Noureen A, Timmer-Bosscha H, Schäfer G, Walenkamp A, Perner S, Beatovic A, Moisse M, Plattner C, Krogsdam A, Haybaeck J, Sopper S, Thaler S, Keller MA, Klocker H, Trajanoski Z, Wolf D, Pircher A. Mol Cancer. 2022 Jun 18;21(1):132. doi: 10.1186/s12943-022-01597-7. PMID: 35717322; PMCID: PMC9206324. Full text

Abstract

Background: Crosstalk between neoplastic and stromal cells fosters prostate cancer (PCa) progression and dissemination. Insight in cell-to-cell communication networks provides new therapeutic avenues to mold processes that contribute to PCa tumor microenvironment (TME) alterations. Here we performed a detailed characterization of PCa tumor endothelial cells (TEC) to delineate intercellular crosstalk between TEC and the PCa TME.

Methods: TEC isolated from 67 fresh radical prostatectomy (RP) specimens underwent multi-omic ex vivo characterization as well as orthogonal validation of both TEC functions and key markers by immunohistochemistry (IHC) and immunofluorescence (IF). To identify cell-cell interaction targets in TEC, we performed single-cell RNA sequencing (scRNA-seq) in four PCa patients who underwent a RP to catalogue cellular TME composition. Targets were cross-validated using IHC, publicly available datasets, cell culture experiments as well as a PCa xenograft mouse model.

Results: Compared to adjacent normal endothelial cells (NEC) bulk RNA-seq analysis revealed upregulation of genes associated with tumor vasculature, collagen modification and extracellular matrix remodeling in TEC. PTGIR, PLAC9, CXCL12 and VDR were identified as TEC markers and confirmed by IF and IHC in an independent patient cohort. By scRNA-seq we identified 27 cell (sub)types, including endothelial cells (EC) with arterial, venous and immature signatures, as well as angiogenic tip EC. A focused molecular analysis revealed that arterial TEC displayed highest CXCL12 mRNA expression levels when compared to all other TME cell (sub)populations and showed a negative prognostic role. Receptor-ligand interaction analysis predicted interactions between arterial TEC derived CXCL12 and its cognate receptor CXCR4 on angiogenic tip EC. CXCL12 was in vitro and in vivo validated as actionable TEC target by highlighting the vessel number- and density- reducing activity of the CXCR4-inhibitor AMD3100 in murine PCa as well as by inhibition of TEC proliferation and migration in vitro.

Conclusions: Overall, our comprehensive analysis identified novel PCa TEC targets and highlights CXCR4/CXCL12 interaction as a potential novel target to interfere with tumor angiogenesis in PCa.

Procedure of the EZ4U colorimetric test

The cell proliferation Assay kit EZ4U is  non-toxic and non-radioactive that is based on the reduction of tetrazolium salt to colored formazan in living cells.  Thus, the assay discriminates between living and dead cells since the process requires functional mitochondria.  The assay is very easy to perform – add EZ4U reagent directly to the cells cultured, incubate and detect absorbance of the supernatant by a reader. The assay has been applied in numerous cells as reported in more than 225 publications  e.g.

Fasting improves therapeutic response in hepatocellular carcinoma through p53-dependent metabolic synergism. Krstic J  et al., Sci Adv. 2022. 21;8(3):eabh2635.

“Cell viability was analyzed using EZ4U assay (Biomedica Immunoassays) according to the manufacturer’s instructions. Briefly, at the end of treatment, the medium was replaced with fresh GM (200 μl per well) and 20 μl per well of EZ4U working solution. After 2 hours of incubation at 37°C, the absorbance was measured at 492 nm with a reference wavelength of 620 nm (Spark 10M multimode microplate reader).”

Syndecan-4 Is a Key Facilitator of the SARS-CoV-2 Delta Variant’s Superior Transmission. Hudák A et al.,  Int J Mol Sci. 2022.  12;23(2):796.

“Cell Viability Measurements: The effect of the applied spike proteins on cell viability was assessed with the EZ4U cell proliferation assay (Biomedica Gmbh, Vienna, Austria, cat. no. BI-5000), according to the instructions of the manufacturer. Absorbance was measured with a BioTek Cytation 3 multimode microplate reader.”

Method

• Water-soluble tetrazolium compound penetrates cell membrane
• In mitochondria reduction takes place and results in intense colored formazan which is water-soluble
• The reagent Formazan is secreted into culture medium and measured with an  ELISA reader at 450 nm or 492 nm

Protocol

• Sample type –  culture medium
• Sample size –  200 µl / test
• Detection limit – depends on respective cell line. Some xamples are shown in the product insert
• Incubation time – between 2-5 hours

 
What does the EZ4U kit include?

The reagents supplied per kit are sufficient for  testing 10 x 96 well microtiter plates and contain:

• Substrate – lyophilized 10 vials
• Activator solution – ready to use 1 x 30 ml

Examples of cell types used in the EZ4U assay

– Endothelial cells (primary isolated from biopsies or cell-lines)
– Peripheral blood mononuclear cells (PBMCsec)
– HeLa (human cervical cancer) and HEK293A (human embryonic kidney) cell lines 
– Human aortic smooth muscle cells (HAoSMCs)
– Huh7 (human liver carcinoma, CLS Cell Lines)
– Colon carcinoma cell models SW480
– CLC CTC cell lines (BHGc7, BHGc10, BHGc16, BHGc26 and UHGc5)
– CRC cell line HT29, DLD-1, HCT116,primary CRC cell line CG08
– HepG2 and Huh6 clone 5 cell lines
– Primary human skin cells – keratinocytes and fibroblasts
– Bovine mammary epithelial cells (MAC-T)
– Human primary chondrocytes
– HL-60 leukemia cells
– A172 and T98G glioblastoma cells
– Clonal preosteoblastic cell line MC3T3-E1-derived from newborn mouse calvaria
– Osteocyte-like MLO-Y4 cell line
– Preosteoclastic, macrophage-like RAW 264.7 cell line
– Primary HUVECs – human vascular endothelial cells
– MCF-7, T47D and MDA-MB-231 breast cancer cell lines
and many more..

 

Baseline Ang-2 plasma levels are an independent prognostic biomarker in refractory metastatic colorectal cancer

Chemorefractory is a term that is used to describe a cancer that does not respond to chemotherapy. Currently, no biomarkers are available to predict the efficacy of chemotherapy in chemorefractory metastatic colorectal cancer. Researchers from Italy have shown that circulating biomarker levels of Angiopoietin-2 (ANG2)  increases early and predicts outcome with regorafenib but not with trifluridine/tipiracil treatment.  Thus, baseline Ang-2 plasma levels are an independent prognostic biomarker in chemorefractory metastatic colorectal cancer. Read more: Early modulation of Angiopoietin-2 plasma levels predicts benefit from regorafenib in patients with metastatic colorectal cancer

Easy measurement of Angiopoietin-2 (ANG-2) in blood samples with only 20µl sample volume.

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Related publications

Early modulation of Angiopoietin-2 plasma levels predicts benefit from regorafenib in patients with metastatic colorectal cancer.

Antoniotti C, Marmorino F, Boccaccino A, Martini S, Antista M, Rossini D, Zuco V, Prisciandaro M, Conca V, Zucchelli G, Borelli B, Cosentino P, Germani MM, Bosco MF, Carullo M, Vetere G, Moretto R, Giordano M, Masi G, Pietrantonio F, Zaffaroni N, Cremolini C. Eur J Cancer. 2022. 165:116-124. doi: 10.1016/j.ejca.2022.01.025. PMID: 35231767.

Abstract

Background: No biomarkers are currently available to predict the efficacy of trifluridine/tipiracil (FTD/TPI) in chemorefractory metastatic colorectal cancer. The multicohort REGOLAND study aims at exploring and validating circulating markers potentially able to predict benefit from regorafenib in this setting. Material and methods: In the retrospective ‘regorafenib exploratory cohort’, including 105 patients treated with regorafenib, baseline (d1) plasma levels of angiogenesis-related biomarkers and their early modulation after 15 days (d15) of treatment were investigated for correlation with clinical outcome. Based on a pre-specified statistical hypothesis, main retrospective findings were prospectively challenged in the ‘regorafenib validation cohort’, including 100 patients treated with regorafenib. Prospectively validated putative biomarkers were then assessed in the control ‘FTD/TPI cohort’, including 93 patients treated with FTD/TPI. Results: In the ‘regorafenib exploratory cohort’, the early (d15) increase of Angiopoietin-2 (Ang-2) was associated with longer progression-free survival (HR:0.57 [95%CI:0.38-0.88], P = 0.004) and a trend towards longer OS (HR:0.74 [95%CI:0.48-1.14], P = 0.165), than the early decrease. Similar results were prospectively confirmed in the ‘regorafenib validation cohort’ (HR for progression-free survival:0.72 [95%CI:0.48-1.08], P = 0.095; HR for OS:0.77 [95%CI:0.51-1.16], P = 0.204). No predictive impact was shown for the early modulation of Ang-2 in the ‘FTD/TPI cohort’. High baseline Ang-2 levels predict poor prognosis in all the investigated cohorts, independently of other clinical prognostic variables. Conclusions: The early modulation of circulating Ang-2 predicts the efficacy of regorafenib. Baseline Ang-2 plasma levels are an independent prognostic biomarker in chemorefractory metastatic colorectal cancer.

Angiopoietin-2 as a Prognostic Factor in Patients with Incurable Stage IV Colorectal Cancer.

Munakata S, Ueyama T, Ishihara H, Komiyama H, Tsukamoto R, Kawai M, Takahashi M, Kojima Y, Tomiki Y, Sakamoto K. J Gastrointest Cancer. 2021.52(1):237-242. PMID: 32166589.

 

 

UK study suggests protective effect of Vitamin D supplementation on bone metabolism

Vitamin D plays an important role in bone mineralization. It helps the body to effectively absorb calcium which is essential to good bone health. The most common way for the body to produce vitamin D is through sunlight. As we get older the natural production of vitamin D decreases. Thus, elderly people either need to take vitamin D supplements or focus on eating foods that contain vitamin D. In a recent study, UK researchers investigated the effect of vitamin D supplementation in older people for 12 months. They looked into changes in markers of bone metabolism, bone mineral density, and bone mineral content . The results suggest a protective effect of supplementation on bone metabolism. Read more: Vitamin D Supplementation for 12 Months in Older Adults Alters Regulators of Bone Metabolism but Does Not Change Wnt Signaling Pathway Markers.

 

Biomedica offers a wide range of high quality ELISA Kits for bone markers.

The following Wnt signaling and regulatory bone markers were measured in this study:

Sclerostin, OPG, RANKL, DKK-1, FGF23 intact, FGF23 (C-terminal)

 

Related publications

Vitamin D Supplementation for 12 Months in Older Adults Alters Regulators of Bone Metabolism but Does Not Change Wnt Signaling Pathway Markers.
Christodoulou M, Aspray TJ, Piec I, Washbourne C, Tang JC, Fraser WD, Schoenmakers I; VDOP Trial Group. JBMR Plus. 2022 Mar 24;6(5):e10619. doi: 10.1002/jbm4.10619. PMID: 35509637; PMCID: PMC9059470.

Abstract
Vitamin D status and supplementation regulates bone metabolism and may modulate Wnt signaling. We studied the response of hormonal regulators of bone metabolism, markers of Wnt signaling and bone turnover and bone mineral density (BMD) and bone mineral content (BMC) in a randomized vitamin D intervention trial (12,000 IU, 24,000 IU, 48,000 IU/mo for 1 year; men and women aged >70 years; n = 379; ISRCTN35648481). Associations with total and free 25(OH)D concentrations were analyzed by linear regression. Baseline vitamin D status was (mean ± SD) 25(OH)D: 40.0 ± 20.1 nmol/L. Supplementation dose-dependently increased total and free 25(OH)D concentrations and decreased plasma phosphate and parathyroid hormone (PTH) (all p < 0.05). The procollagen 1 intact N-terminal (PINP)/C-terminal telopeptide (CTX) ratio, C-terminal fibroblast growth factor-23 (cFGF23), and intact FGF23 (iFGF23) significantly increased with no between-group differences, whereas Klotho was unchanged. 1,25(OH)2D and PINP significantly increased in the 24 IU and 48,000 IU groups. Sclerostin (SOST), osteoprotegerin (OPG), receptor activator of NF-κB ligand (RANKL), BMD, BMC, and CTX remained unchanged. Subgroup analyses with baseline 25(OH)D <25 nmol/L (n = 94) provided similar results. Baseline total and free 25(OH)D concentrations were positively associated with 1,25(OH)2D, 24,25(OH)2D (p < 0.001), vitamin D binding protein (DBP) (p < 0.05), BMD, and BMC (p < 0.05). Associations with PTH (p <0.001), cFGF23 (p < 0.01), and BAP (p < 0.05) were negative. After supplementation, total and free 25(OH)D concentrations remained positively associated only with 24,25(OH)2D (p < 0.001) and DBP (p < 0.001) and negatively with estimated glomerular filtration rate (eGFR) (p < 0.01). PTH and SOST were significantly associated only with free 25(OH)D. There were no significant relationships with BMD and BMC after supplementation. The decrease in PTH and increase in PINP/CTX ratio suggest a protective effect of supplementation on bone metabolism, although no significant effect on BMD or pronounced changes in regulators of Wnt signaling were found. The increase in FGF23 warrants caution because of its negative association with skeletal and cardiovascular health. Associations of total and free 25(OH)D with biomarkers were similar and known positive associations between vitamin D status and BMD were confirmed. The change in associations after supplementation might suggest a threshold effect. © 2022 The Authors. JBMR Plus published by Wiley Periodicals LLC on behalf of American Society for Bone and Mineral Research.

The role of vitamin D in maintaining bone health in older people.
Hill TR, Aspray TJ. Ther Adv Musculoskelet Dis. 2017 Apr;9(4):89-95. doi: 10.1177/1759720X17692502. Epub 2017 Feb 14. PMID: 28382112; PMCID: PMC5367643.

Abstract
This review summarises aspects of vitamin D metabolism, the consequences of vitamin D deficiency, and the impact of vitamin D supplementation on musculoskeletal health in older age. With age, changes in vitamin D exposure, cutaneous vitamin D synthesis and behavioural factors (including physical activity, diet and sun exposure) are compounded by changes in calcium and vitamin D pathophysiology with altered calcium absorption, decreased 25-OH vitamin D [25(OH)D] hydroxylation, lower renal fractional calcium reabsorption and a rise in parathyroid hormone. Hypovitaminosis D is common and associated with a risk of osteomalacia, particularly in older adults, where rates of vitamin D deficiency range from 10-66%, depending on the threshold of circulating 25(OH)D used, population studied and season. The relationship between vitamin D status and osteoporosis is less clear. While circulating 25(OH)D has a linear relationship with bone mineral density (BMD) in some epidemiological studies, this is not consistent across all racial groups. The results of randomized controlled trials of vitamin D supplementation on BMD are also inconsistent, and some studies may be less relevant to the older population, as, for example, half of participants in the most robust meta-analysis were aged under 60 years. The impact on BMD of treating vitamin D deficiency (and osteomalacia) is also rarely considered in such intervention studies. When considering osteoporosis, fracture risk is our main concern, but vitamin D therapy has no consistent fracture-prevention effect, except in studies where calcium is coprescribed (particularly in frail populations living in care homes). As a J-shaped effect on falls and fracture risk is becoming evident with vitamin D interventions, we should target those at greatest risk who may benefit from vitamin D supplementation to decrease falls and fractures, although the optimum dose is still unclear.

The health effects of vitamin D supplementation: evidence from human studies.
Bouillon R, Manousaki D, Rosen C, Trajanoska K, Rivadeneira F, Richards JB. Nat Rev Endocrinol. 2022 Feb;18(2):96-110. doi: 10.1038/s41574-021-00593-z. Epub 2021 Nov 23. PMID: 34815552; PMCID: PMC8609267.

Abstract
Vitamin D supplementation can prevent and cure nutritional rickets in infants and children. Preclinical and observational data suggest that the vitamin D endocrine system has a wide spectrum of skeletal and extra-skeletal activities. There is consensus that severe vitamin D deficiency (serum 25-hydroxyvitamin D (25OHD) concentration <30 nmol/l) should be corrected, whereas most guidelines recommend serum 25OHD concentrations of >50 nmol/l for optimal bone health in older adults. However, the causal link between vitamin D and many extra-skeletal outcomes remains unclear. The VITAL, ViDA and D2d randomized clinical trials (combined number of participants >30,000) indicated that vitamin D supplementation of vitamin D-replete adults (baseline serum 25OHD >50 nmol/l) does not prevent cancer, cardiovascular events, falls or progression to type 2 diabetes mellitus. Post hoc analysis has suggested some extra-skeletal benefits for individuals with vitamin D deficiency. Over 60 Mendelian randomization studies, designed to minimize bias from confounding, have evaluated the consequences of lifelong genetically lowered serum 25OHD concentrations on various outcomes and most studies have found null effects. Four Mendelian randomization studies found an increased risk of multiple sclerosis in individuals with genetically lowered serum 25OHD concentrations. In conclusion, supplementation of vitamin D-replete individuals does not provide demonstrable health benefits. This conclusion does not contradict older guidelines that severe vitamin D deficiency should be prevented or corrected.

Vitamin D: Production, Metabolism and Mechanisms of Action.
Bikle DD. 2021. Feingold KR, Anawalt B, Boyce A, Chrousos G, de Herder WW, Dhatariya K, Dungan K, Hershman JM, Hofland J, Kalra S, Kaltsas G, Koch C, Kopp P, Korbonits M, Kovacs CS, Kuohung W, Laferrère B, Levy M, McGee EA, McLachlan R, Morley JE, New M, Purnell J, Sahay R, Singer F, Sperling MA, Stratakis CA, Trence DL, Wilson DP, editors. Endotext [Internet]. South Dartmouth (MA): MDText.com, Inc.; 2000–. PMID: 25905172.

Excerpt
Vitamin D production in the skin under the influence of sunlight (UVB) is maximized at levels of sunlight exposure that do not burn the skin. Further metabolism of vitamin D to its major circulating form (25(OH)D) and hormonal form (1,25(OH)2D) takes place in the liver and kidney, respectively, but also in other tissues where the 1,25(OH)2D produced serves a paracrine/autocrine function: examples include the skin, cells of the immune system, parathyroid gland, intestinal epithelium, prostate, and breast. Parathyroid hormone, FGF23, calcium and phosphate are the major regulators of the renal 1-hydroxylase (CYP27B1, the enzyme producing 1,25(OH)2D); regulation of the extra renal 1-hydroxylase differs from that in the kidney and involves cytokines. The major enzyme that catabolizes 25(OH)D and 1,25(OH)2D is the 24-hydroxylase; like the 1-hydroxylase it is tightly controlled in the kidney in a manner opposite to that of the 1-hydroxylase, but like the 1-hydroxylase it is widespread in other tissues where its regulation is different from that of the kidney. Vitamin D and its metabolites are carried in the blood bound to vitamin D binding protein (DBP) and albumin–for most tissues it is the free (i.e., unbound) metabolite that enters the cell; however, DBP bound metabolites can enter some cells such as the kidney and parathyroid gland through a megalin/cubilin mechanism. Most but not all actions of 1,25(OH)2D are mediated by the vitamin D receptor (VDR). VDR is a transcription factor that partners with other transcription factors such as retinoid X receptor that when bound to 1,25(OH)2D regulates gene transcription either positively or negatively depending on other cofactors to which it binds or interacts. The VDR is found in most cells, not just those involved with bone and mineral homeostasis (i.e., bone, gut, kidney) resulting in wide spread actions of 1,25(OH)2D on most physiologic and pathologic processes. Animal studies indicate that vitamin D has beneficial effects on various cancers, blood pressure, heart disease, immunologic disorders, but these non-skeletal effects have been difficult to prove in humans in randomized controlled trials. Analogs of 1,25(OH)2D are being developed to achieve specificity for non-skeletal target tissues such as the parathyroid gland and cancers to avoid the hypercalcemia resulting from 1,25(OH)2D itself. The level of vitamin D intake and achieved serum levels of 25(OH)D that are optimal and safe for skeletal health and the non-skeletal actions remain controversial, but are likely between an intake of 800-2000IU vitamin D in the diet and 20-50ng/ml 25(OH)D in the blood.

Sleeve gastrectomy might improve the “natriuretic handicap” of severely obese individuals

NT-proBNP serum levels increase with surgical weight loss interventions: Natriuretic peptides, including BNP hormone (B-type natriuretic peptide) and its amino-terminal counterpart NT-proBNP  play a crucial role in the regulation of cardiovascular and energy homeostasis. They decrease blood pressure and increase the degradation of lipids and energy expenditure in fat tissue. Furthermore, both hormones are used as biomarkers in clinical practice in the diagnosis of heart failure.

Several studies have demonstrated that obesity is associated with low circulating levels of NT-proBNP concentrations, a condition known as “natriuretic handicap” * .  In the following publication, researchers have investigated the impact of two surgical weight loss interventions on NT-proBNP serum concentrations in obese individuals without clinical signs of heart failure.  Their data support the idea that the “natriuretic handicap * ” of obese individuals can be improved with surgically-induced weight loss, especially sleeve gastrectomy.

Effect of various weight loss interventions on serum NT-proBNP concentration in severe obese subjects without clinical manifest heart failure. Hollstein T et al. 2021.

*condition of decreased BNP levels in obese patients has been termed “the natriuretic handicap ” ( B-type natriuretic peptide and obesity in heart failure: a mysterious but important association in clinical practice. Reinmann M and P Meyer. 2020.)

⇒ NT-proBNP serum concentrations were measured with the Biomedica NT-proBNP ELISA (cat.no. SK-1204)

NT-proBNP serum levels increase with surgical weight loss interventions – 

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Developed & manufactured by Biomedica – right in the heart of Europe

 

Effect of various weight loss interventions on serum NT-proBNP concentration in severe obese subjects without clinical manifest heart failure.

Hollstein T, Schlicht K, Krause L, Hagen S, Rohmann N, Schulte DM, Türk K, Beckmann A, Ahrens M, Franke A, Schreiber S, Becker T, Beckmann J, Laudes M. Sci Rep. 2021. 12;11(1):10096. doi: 10.1038/s41598-021-89426-7. PMID: 33980890; PMCID: PMC8115663.

Abstract

The B-type natriuretic peptide (BNP) hormone plays a crucial role in the regulation of cardiovascular and energy homeostasis. Obesity is associated with low circulating levels of BNP, a condition known as “natriuretic handicap.” Recent evidences suggest an altered expression of BNP receptors-both the signaling natriuretic peptide receptors (NPR)-A and the clearance NPR-C receptor-in adipose tissue (AT) as one of the putative causes of natriuretic handicap.

The current study aims at clarifying the molecular mechanisms behind the natriuretic handicap, focusing on NPR modulation in the AT of obese and control subjects.

The study enrolled 34 obese and 20 control subjects undergoing bariatric or abdominal surgery, respectively. The main clinical and biochemical parameters, including circulating BNP, were assessed. In visceral (VAT) and subcutaneous AT (SAT) samples, collected during surgery, the adipocytes and stromal vascular fraction (SVF) expression of NPR-A and NPR-C and the SVF secretion of interleukin 6 (IL-6) were determined. Both VAT and SAT from obese patients expressed a lower NPR-A/NPR-C ratio in adipocytes and the SVF secreted a higher level of IL-6, compared with the controls. Moreover, NPR-A/NPR-C ratio expressed by VAT and SAT adipocytes negatively correlated with body mass index, insulin, the Homeostasis Model Assessment of Insulin resistance, and IL-6 secreted by SVF, and the expression of the clearance receptor NPR-C, in both the VAT and SAT adipocytes, showed a negative correlation with circulating BNP.

Overall, insulin resistance/hyperinsulinemia and AT inflammation (ie, high level of IL-6) are the major determinants of the lower NPR-A/NPR-C ratio in adipocytes, thus contributing to the natriuretic handicap in obese subjects.

 

RELATED PUBLICATIONS

 Modulation of natriuretic peptide receptors in human adipose tissue: molecular mechanisms behind the “natriuretic handicap” in morbidly obese patients.

Gentili A et al., . Transl Res. 2017. 186:52-61. doi: 10.1016/j.trsl.2017.06.001. PMID: 28651075.

Abstract

The B-type natriuretic peptide (BNP) hormone plays a crucial role in the regulation of cardiovascular and energy homeostasis. Obesity is associated with low circulating levels of BNP, a condition known as “natriuretic handicap.” Recent evidences suggest an altered expression of BNP receptors-both the signaling natriuretic peptide receptors (NPR)-A and the clearance NPR-C receptor-in adipose tissue (AT) as one of the putative causes of natriuretic handicap.

The current study aims at clarifying the molecular mechanisms behind the natriuretic handicap, focusing on NPR modulation in the AT of obese and control subjects.

The study enrolled 34 obese and 20 control subjects undergoing bariatric or abdominal surgery, respectively. The main clinical and biochemical parameters, including circulating BNP, were assessed. In visceral (VAT) and subcutaneous AT (SAT) samples, collected during surgery, the adipocytes and stromal vascular fraction (SVF) expression of NPR-A and NPR-C and the SVF secretion of interleukin 6 (IL-6) were determined. Both VAT and SAT from obese patients expressed a lower NPR-A/NPR-C ratio in adipocytes and the SVF secreted a higher level of IL-6, compared with the controls. Moreover, NPR-A/NPR-C ratio expressed by VAT and SAT adipocytes negatively correlated with body mass index, insulin, the Homeostasis Model Assessment of Insulin resistance, and IL-6 secreted by SVF, and the expression of the clearance receptor NPR-C, in both the VAT and SAT adipocytes, showed a negative correlation with circulating BNP.

Overall, insulin resistance/hyperinsulinemia and AT inflammation (ie, high level of IL-6) are the major determinants of the lower NPR-A/NPR-C ratio in adipocytes, thus contributing to the natriuretic handicap in obese subjects.

 

Obesity and Serial NT-proBNP Levels in Guided Medical Therapy for Heart Failure With Reduced Ejection Fraction: Insights From the GUIDE-IT Trial.

Parcha V, Patel N, Kalra R, Suri SS, Arora G, Wang TJ, Arora P. J Am Heart Assoc. 2021 Apr 6;10(7):e018689. doi: 10.1161/JAHA.120.018689. PMID: 33754794; PMCID: PMC8174357.

Abstract

Background Obese patients have lower NT-proBNP (N-terminal pro-B-type natriuretic peptide) levels.

The prognostic implications of achieving NT-proBNP levels ≤1000 pg/mL in obese patients with heart failure (HF) receiving biomarker-guided therapy are not completely known.

We evaluated the prognostic implications of obesity and having NT-proBNP levels (≤1000 pg/mL) in the GUIDE-IT (Guiding Evidence-Based Therapy Using Biomarker-Intensified Treatment in HF) trial participants. Methods and Results The risk of adverse cardiovascular events (HF hospitalization or cardiovascular mortality) was assessed using multivariable-adjusted Cox proportional hazard models based on having NT-proBNP ≤1000 pg/mL (taken as a time-varying covariate), stratified by obesity status. The study outcome was also assessed on the basis of the body mass index at baseline. The predictive ability of NT-proBNP for adverse cardiovascular events was assessed using the likelihood ratio test. Compared with nonobese patients, obese patients were mostly younger, Black race, and more likely to be women. NT-proBNP levels were 59.0% (95% CI, 39.5%-83.5%) lower among obese individuals. The risk of adverse cardiovascular events was lower in obese (hazard ratio [HR], 0.48; 95% CI, 0.29-0.59) and nonobese (HR, 0.32; 95% CI, 0.19-0.57) patients with HF who had NT-proBNP levels ≤1000 pg/mL, compared with those who did not. There was no interaction between obesity and having NT-proBNP ≤1000 pg/mL on the study outcome (P>0.10). Obese patients had a greater risk of developing adverse cardiovascular events (HR, 1.39; 95% CI, 1.01-1.90) compared with nonobese patients. NT-proBNP was the strongest predictor of adverse cardiovascular event risk in both obese and nonobese patients.

Conclusions On-treatment NT-proBNP level ≤1000 pg/mL has favorable prognostic implications, irrespective of obesity status. NT-proBNP levels were the strongest predictor of cardiovascular events in both obese and nonobese individuals in this trial.

Registration URL: https://www.clinicaltrials.gov; Unique identifier: NCT01685840.

BIOMEDICA Quality CYTOKINE KITS for affordable & efficient research

My name is Lan Chi. I am part of the Biomedica assay quality management team. All our ELISA kits undergo a stringent quality control process. This ensures reliable and consistent results. We are located right in the heart of Europe.  I am happy to introduce our new human Cytokine ELISA Kits  – developed & manufactured by Biomedica 

BIOMEDICA Quality CYTOKINE KITS for affordable & efficient research.

 

IL-6 (Interleukin-6)                         high sensitivity – detectable levels

VEGF                                                       low sample volume – 10µl

ANGIOPOIETIN-2                          optimized assay range – no predilution

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WE VALIDATE OUR KITS ACCORDING to INTERNATIONAL QUALITY STANDARDS 

 

Biomedica ´s ELISA assay kits are validated according to international quality guidelines (ICH, EMEA, FDA).  You can find the assay validation data on the respective product pages of our website.  

DILUTION LINEARITY (PARALLELISM): CONSISTENT RECOVERY IN DILUTED SAMPLES

Our validation data include the following experiments: e.g. parallelism (dilution of samples containing elevated endogenous levels of the protein of interest) and dilution linearity (dilution of samples containing levels of endogenous and recombinant protein of interest).

WITHIN-RUN AND IN-BETWEEN RUN PRECISION: PRECISE & REPRODUCIBLE RESULTS WITHIN AND ACROSS LOTS

Our lab team also validates our assays for precision and accuracy. The precision of our assays is determined by measuring  samples of known concentrations a number of times with one kit lot (within-run precision) and with two different kit lots (in-between run precision). 

Finally, we check the stability of the analyte in the respective sample matrix.

STABILITY – Sample stability and stability of kit components

We take measures to ensure the stability of all component substances that are included in the kits. Accordingly, the stability of all assay components is tested during kit development. Furthermore, we expose real samples to several freeze-thaw cycles and also leave samples at room temperature for at least 24 hours. Hence, with these experiments we are able to determine sample stability after temperature stress. 

Related information:

Our Biomedica human Angiopoietin-2 ELISA  (cat.no. BI-ANG2) has been cited in the following publication:

Angiopoietin-2 predicts all-cause mortality in male but not female end-stage kidney disease patients on hemodialysis.

Chu C et al., Nephrol Dial Transplant. 2022 ;  23;37(7):1348-1356.

Particularly, a detailed description of Biomedica´s Angiopoietin-2 ELISA is shown in the following poster .  

Check out the validation data of our human IL-6 ELISA

We offer tailored analytical testing services for the measurement of your unique samples.  

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For ELISA  & Luminex Assays, microRNA Analysis, and Glycan Profiling.

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Check out our ELISA and Luminex Workflow chart and our microRNA Service Details :

Biomedica offers a broad range of high quality RNA services performed by experts according to GLP standards. These services include:

• RNA extraction: Biofluids including serum, plasma, and extracellular vesicles. Cells and tissues – the quality control of total RNA is carried out utilizing bioanalyser chips. 
• Next-generation sequencing- NGS.
• RT-qPCR.
• We also analyse cell-type specific microRNA/mRNA  in complex tissues and offer custom analysis of microRNA signatures.
• Finally, we carry out the analysis of established microRNA signatures to predict fracture risk, platelet function, and toxicity.
• Biomedica and TAmiRNA 

• Biomedica Service Measurements  – reliable and flexible 

  We can measure any kind of selected biomarker by ELISA.

  We also offer our own propriety Biomedica biomarker ELISA kits – that are developed and manufactured in our laboratories. 
  For more information on our product lines, see our product list.

• Our Service Measurement Applications include 

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  • Custom applications

Testicular Cancer – RANKL inhibition suppresses tumor growth: Testicular cancer is the most commonly diagnosed malignancy in young males. A new study features the use of the Biomedica RANKL ELISA investigating the effects of RANKL (receptor activator of nuclear factor- κB ligand) inhibition in patients with testicular cancer. Despite its high cure rate, patients suffer from serious adverse effects to chemotherapy. Investigating alternative treatments, the scientists demonstrated that the RANKL inhibitor Denosumamb, used to treat osteoporosis, has tumor suppressive properties. As RANKL signaling was recently identified in the testis regulating male reproductive function, an open question remains if RANKL is responsible for the malignant transformation to invasive tumors.

Testicular Cancer – RANKL inhibition suppresses tumor growth

 

RANKL regulates testicular cancer growth and Denosumab treatment has suppressive effects on GCNIS and advanced seminoma.                                                                                                                                        

Andreassen CH, Lorenzen M, Nielsen JE, Kafai Yahyavi S, Toft BG, Ingerslev LR, Clemmensen C, Rasmussen LJ, Bokemeyer C, Juul A, Jørgensen A, Blomberg Jensen M. Br J Cancer. 2022 Apr 13. doi: 10.1038/s41416-022-01810-w. Epub ahead of print. PMID: 35418213.

 

Abstract

Background: Testicular germ cell tumours (TGCTs) have a high sensitivity to chemotherapy and a high cure rate, although with serious adverse effects. In the search for tumour suppressive drugs, the RANKL inhibitor Denosumab, used to treat osteoporosis, came up as a candidate since RANKL signalling was recently identified in the testis.

Methods: Expression of RANKL, RANK and OPG, and the effects of RANKL inhibition were investigated in human TGCTs, TGCT-derived cell-lines, and TGCT-xenograft models. Serum RANKL was measured in TGCT-patients.

Results: RANKL, RANK, and OPG were expressed in germ cell neoplasia in situ (GCNIS), TGCTs, and TGCT-derived cell lines. RANKL-inhibition reduced proliferation of seminoma-derived TCam-2 cells, but had no effect on embryonal carcinoma-derived NTera2 cells. Pretreatment with Denosumab did not augment the effect of cisplatin in vitro. However, inhibition of RANKL in vivo reduced tumour growth exclusively in the TCam-2-xenograft model and Denosumab-treatment decreased proliferation in human GCNIS cultures. In TGCT-patients serum RANKL had no prognostic value.

Conclusions: This study shows that the RANKL signalling system is expressed in GCNIS and seminoma where RANKL inhibition suppresses tumour growth in vitro and in vivo. Future studies are needed to determine whether RANKL is important for the malignant transformation or transition from GCNIS to invasive tumours.

 

FREE soluble RANKL HS ELISA | BI-20462

 

Biomedica offers an ELISA assay kit for the reliable detection of  free soluble uncomplexed RANKL in human serum samples: free soluble RANKL ELISA (cat. no. BI-20462)

 Widely cited in clinical studies – 290+ publications

• Reliable – fully validated according to international guidelines
• High sensitivity – measurable concentrations in healthy subjects
• HIGH quality guaranteed  – developed & produced by Biomedica

 

Related products

Osteoprotegerin (OPG) ELISA  , DKK-1 ELISA ,  Sclerostin ELISA ,

IL-6 ELISA  ,  VEGF ELISA ,  Angiopoietin-2 ELISA

 

Related publications

RANKL regulates male reproductive function.

Blomberg Jensen M, Andreassen CH, Jørgensen A, Nielsen JE, Juel Mortensen L, Boisen IM, et al. Nat Commun. 2021;12:1–15. doi: 10.1038/s41467-021-22734-8. PMID: 33893301; PMCID: PMC8065035.

Abstract

Infertile men have few treatment options. Here, we demonstrate that the transmembrane receptor activator of NF-kB ligand (RANKL) signaling system is active in mouse and human testis. RANKL is highly expressed in Sertoli cells and signals through RANK, expressed in most germ cells, whereas the RANKL-inhibitor osteoprotegerin (OPG) is expressed in germ and peritubular cells. OPG treatment increases wild-type mouse sperm counts, and mice with global or Sertoli-specific genetic suppression of Rankl have increased male fertility and sperm counts. Moreover, RANKL levels in seminal fluid are high and distinguishes normal from infertile men with higher specificity than total sperm count. In infertile men, one dose of Denosumab decreases RANKL seminal fluid concentration and increases serum Inhibin-B and anti-Müllerian-hormone levels, but semen quality only in a subgroup. This translational study suggests that RANKL is a regulator of male reproductive function, however, predictive biomarkers for treatment-outcome requires further investigation in placebo-controlled studies.

The RANK-RANKL axis: an opportunity for drug repurposing in cancer?

Peters S, Clézardin P, Márquez-Rodas I, Niepel D, Gedye C. Clin Transl Oncol. 2019;21:977–91. doi: 10.1007/s12094-018-02023-5. Epub 2019 Jan 17. PMID: 30656607.

Abstract

Drug repurposing offers advantages over traditional drug development in terms of cost, speed and improved patient outcomes. The receptor activator of nuclear factor kappa B (RANK) ligand (RANKL) inhibitor denosumab is approved for the prevention of skeletal-related events in patients with advanced malignancies involving bone, including solid tumours and multiple myeloma. Following improved understanding of the role of RANK/RANKL in cancer biology, denosumab has already been repurposed as a treatment for giant cell tumour of bone. Here, we review the role of RANK/RANKL in tumourigenesis, including effects on tumour initiation, progression and metastasis and consider the impact of RANK/RANKL on tumour immunology and immune evasion. Finally, we look briefly at ongoing trials and future opportunities for therapeutic synergy when combining denosumab with anti-cancer agents such as immune checkpoint inhibitors.

Possible link between FSH and RANKL release from adipocytes in men with impaired gonadal function including Klinefelter syndrome .

Juel Mortensen L, Lorenzen M, Jørgensen N, Andersson A-M, Nielsen JE, Petersen LI, et al.Bone. 2019;123:103–14. doi: 10.1016/j.bone.2019.03.022. Epub 2019 Mar 23. PMID: 30914274.

International Trends in the Incidence of Testicular Cancer: Lessons from 35 Years and 41 Countries.

Gurney JK, Florio AA, Znaor A, Ferlay J, Laversanne M, Sarfati D, Bray F, McGlynn KA. Eur Urol. 2019 Nov;76(5):615-623. doi: 10.1016/j.eururo.2019.07.002. Epub 2019 Jul 17. PMID: 31324498; PMCID: PMC8653517.

Development of a 3D-printed testicular cancer model for testicular examination education.

Power RJ, Hearn J, Gillis CJ, Harvey D, French C, Organ M. Can Urol Assoc J. 2021 Apr;15(4):E221-E226. doi: 10.5489/cuaj.6675. PMID: 33007179; PMCID: PMC8021429.

 

How to do a Testicular Self Exam . Testicular Cancer Society

Circulating Angiopoietin-2 (ANG2) increases with PAP sleep apnea treatment: Obstructive sleep apnea (OSA), the most common sleep-related breathing disorder, is becoming increasingly prevalent worldwide due to widespread obesity. It causes a person to repeatedly stop and start breathing during sleep.  Positive airway pressure (PAP) is a well-established treatment for OSA patients, using a machine to pump air under pressure into the airway of the lungs. Though effective, the therapy causes large increases in lung volumes during the night that are potentially deleterious.  A recent study investigated the impact of PAP therapy on various biomarkers related to alveolar epithelial and endothelial injury. The researchers have shown that Angiopoietin-2 (ANG-2), a marker of endothelial injury and cardiovascular disease risk, increases with continuous positive airway pressure therapy . This finding raises concern for a possible adverse impact of PAP therapy on the vascular endothelium.

Circulating-angiopoietin-2-ang2-increases-with-pap-sleep-apnea-treatment: Effect of positive airway pressure therapy of obstructive sleep apnea on circulating Angiopoietin-2.

Gottlieb DJ, Lederer DJ, Kim JS, Tracy RP, Gao S, Redline S, Jelic S. Sleep Med. 2022 May 16;96:119-121. doi: 10.1016/j.sleep.2022.05.007. Epub ahead of print. PMID: 35636149.

Abstract

Background: Obstructive sleep apnea (OSA) has been identified as a possible contributor to interstitial lung disease. While positive airway pressure (PAP) is effective therapy for OSA, it causes large increases in lung volumes during the night that are potentially deleterious, analogous to ventilator-induced lung injury, although this has not been previously studied. The goal of this study was to assess the impact of PAP therapy on four biomarkers of alveolar epithelial and endothelial injury and extracellular matrix remodeling in patients with OSA.

Methods: In 82 patients with moderate to severe OSA who were adherent to PAP therapy, surfactant protein D, osteopontin, angiopoietin-2, and matrix metalloprotease-7 were measured by ELISA in serum samples collected before and 3- to 6-months after initiation of PAP therapy.

Results: An increase in angiopoietin-2 level of 0.28 ng/mL following PAP therapy was observed (p = 0.007). This finding was replicated in an independent sample of OSA patients. No significant change was detected in surfactant protein D, osteopontin, or matrix metalloprotease-7.

Conclusions: This finding raises concern for a possible adverse impact of PAP therapy on vascular endothelium.

Keywords: Angiopoietin-2; Lung injury; Obstructive sleep apnea; Positive airway pressure.

Human Angiopoietin-2 ELISA Kit | BI-ANG2

Angiopoietin-2 can easily be measured with an ELISA assay in serum and plasma samples with only 20 µl of sample volume! The kit incorporates ready to use standards and controls. The assay range is optimized for clinical samples- no sample dilution required!

Features & Benefits of the Biomedica human Angipoietin-2 ELISA

√ Immediate results: no sample dilution required
√ Full validation package – the assay is optimized for clinical samples
√ Kit includes ready to use standards and controls
√ HIGH QUALITY GUARANTEED

Find out more: Angiopoietin-2 ELISA

Circulating-angiopoietin-2 increases-with-pap-sleep-apnea-treatment 

Related publications:

Increased Level of Angiopoietin Like Proteins 4 and 8 in People With Sleep Apnea.

Al-Terki A, Abu-Farha M, AlKhairi I, Cherian PT, Sriraman D, Shyamsundar A, Ali S, Almulla F, Tuomilehto J, Abubaker JA. Front Endocrinol (Lausanne). 2018 Nov 13;9:651. doi: 10.3389/fendo.2018.00651. PMID: 30524367; PMCID: PMC6262344.

Abstract

Objective: Obstructive sleep apnea (OSA) is a sleep disorder caused by the complete or partial obstruction of the upper airways. The worldwide prevalence of OSA is increasing due to its close association with obesity epidemic and multiple health complications, such as hypertension, cardiovascular disease, and Type 2 diabetes. Angiopoietin-like protein (ANGPTL)-4 and ANGPTL8 (betatrophin) have been suggested to play a role in the development of these diseases through their role in regulating the metabolism of plasma lipid molecules. This study was designed to evaluate ANGPTL4 and 8 levels in an OSA group and a control group to clarify the effect of OSA on ANGPTL4 and 8 levels. Methods: In total, 74 subjects were enrolled in this study, including 22 age- and body mass index (BMI)-matched controls with the Apnea Hypopnea Index (AHI) score of <5 events/h and 52 subjects with an AHI score of >5 events/h. Sleep apnea was assessed using a portable sleep test. ANGPTL4 and 8 levels were measured in plasma samples using enzyme-linked immunosorbent assay. Results: Mean AHI score (2.5 ± 1.6) in the control group was significantly lower than that in the OSA group (22.9 ± 17.9; p < 0.0001). Leptin, interleukin-(IL) 6, insulin, and HOMA-IR values were higher in the OSA group than in the control group. ANGPTL8 level was higher in the OSA group (1130.0 ± 108.61 pg/mL) than in the control group (809.39 ± 108.78 pg/mL; p = 0.041). Similarly, ANGPTL4 was higher in the OSA group (179.26 ± 12.89 ng/mL) than in the control group (142.63 ±7.99 ng/mL; p = 0.018). Conclusion: Our findings demonstrate that ANGPTL4 and 8 levels were increased in subjects with OSA, suggesting that the upregulation of these lipid metabolism regulators might play a role in lipid dysregulation observed in people with OSA.

Complement promotes endothelial von Willebrand factor and angiopoietin-2 release in obstructive sleep apnea.

Gao S, Emin M, Thoma T, Pastellas K, Castagna F, Shah R, Jimenez A, Patel N, Wei Y, Jelic S. Sleep. 2021 Apr 9;44(4):zsaa286. doi: 10.1093/sleep/zsaa286. PMID: 33351148; PMCID: PMC8033461.

Abstract

Study objective: Obstructive sleep apnea (OSA) is highly prevalent and triples vascular thromboembolic risk. Intermittent hypoxia (IH) during transient cessation of breathing in OSA impairs endothelial protection against complement. Complement activation stimulates the endothelial release of a pro-thrombotic von Willebrand factor (vWF). We investigated whether increased complement activity in OSA promotes the endothelial release of vWF and pro-inflammatory angiopoietin-2. We further investigated whether improving complement protection with statins reverses these changes.

Methods: Using endothelial cells (ECs) and blood collected from OSA patients (n = 109) and controls (n = 67), we assessed whether altered cellular localization of complement inhibitor CD59 in OSA modulates exocytosis of Weibel-Palade bodies (WPB), secretory granules that store vWF and angiopoietin-2. These interactions were also assessed in vitro in ECs exposed to normoxia or IH with or without recombinant complement C9 and with or without atorvastatin.

Results: Circulating levels of angiopoietin-2 were greater in OSA than controls and levels of vWF cleavage products correlated with OSA severity. In cultured ECs, IH enhanced complement-stimulated angiopoietin-2 and vWF release by reducing EC surface and increasing intracellular expression of complement inhibitor CD59. Intracellular CD59 co-localized with WPB in OSA. IH increased binding of intracellular CD59 to syntaxin-3, which dissociated syntaxin-3 from voltage-sensitive calcium channel Cav1.2, and activated WPB exocytosis in a calcium-dependent manner. Atorvastatin reversed IH-enhanced endothelial release of vWF and angiopoietin-2.

Conclusions: IH promotes the complement-mediated release of vWF and angiopoietin-2, which may contribute to pro-thrombotic and pro-inflammatory conditions in OSA. Statin reversed these effects, suggesting a potential approach to reduce cardiovascular risk in OSA.

Globally, about 1 in 6 deaths is related to cancer. Advances in cancer research have improved the prevention, the detection, and the treatment of cancer. Understanding on how cancers starts, grows and spreads is important for cancer treatment. Biomarkers play a critical role at all stages of the disease and serve as therapeutic targets (1).  

Biomarkers in Cancer Research

NEUROPILIN-1 is a checkpoint target

The biomarker Neuropilin-1 (NRP1) has gained renewed attention as it is implicated in promoting tumor progression. It acts as a co-receptor to VEGF (Vascular Endothelial Growth Factor) and induces angiogenesis, the process of the formation of new blood-vessels (2). NRP-1 is expressed in a variety of cancers including lung, prostate, pancreas or colon carcinoma. In metastatic melanoma, NRP-1 plays a crucial role in melanoma aggressiveness and evidence supports its use as a target for therapies (3) .

More recently, Neuropilin-1 has been identified as a checkpoint target with unique implications for cancer immunology and immunotherapy (4). This review discusses the increasing literature on Neuropilin-1 mediated immune modulation providing a rationale to categorize NRP1 as a key checkpoint in the tumor microenvironment (TME) as well as a promising immunotherapeutic target.

Did you know:  Neuropilin-1 can easily be detected in serum, plasma and in cell culture supernatants using a conventional ELISA kit?

The assay is fully validated for clinical use in human samples but also works in non-human samples. Only 10µl sample is required.

 For more information contact us   at BIOMEDICA

 Neuropilin-1 (NRP1) ELISA assay highlights:

-Highly specific using epitope mapped antibodies
-Detects free and ligand-bound soluble NRP1
-10 µl sample/well

  Literature:

1. Biomarkers and Its Application in Cancer Research. Jiancheng Hu and Qiang You. Biomark Applic 2017: 103. DOI: 10.29011/BMAP-103. 100103.
2. VEGF binding to NRP1 is essential for VEGF stimulation of endothelial cell migration, complex formation between NRP1 and VEGFR2, and signaling via FAK Tyr407 phosphorylation. Herzog B, Pellet-Many C, Britton G, Hartzoulakis B, Zachary IC. Mol Biol Cell. 2011 :;22(15):2766-76. doi: 10.1091/mbc.E09-12-1061. PMID: 21653826.
3. Neuropilin-1 as Therapeutic Target for Malignant Melanoma. Graziani G, Lacal PM. Front Oncol. 2015;5:125. Published 2015 Jun 3. doi:10.3389/fonc.2015.00125.
4. Neuropilin-1: a checkpoint target with unique implications for cancer immunology and immunotherapy. Chuckran CA, Liu C, Bruno TC, Workman CJ, Vignali DA. J Immunother Cancer. 2020 Jul;8(2):e000967. doi: 10.1136/jitc-2020-000967. PMID: 32675311; PMCID: PMC7368550:

Neuropilin-1: a checkpoint target with unique implications for cancer immunology and immunotherapy

Abstract

Checkpoint blockade immunotherapy established a new paradigm in cancer treatment: for certain patients curative treatment requires immune reinvigoration. Despite this monumental advance, only 20%-30% of patients achieve an objective response to standard of care immunotherapy, necessitating the consideration of alternative targets. Optimal strategies will not only stimulate CD8⁺ T cells, but concomitantly modulate immunosuppressive cells in the tumor microenvironment (TME), most notably regulatory T cells (T_reg cells). In this context, the immunoregulatory receptor Neuropilin-1 (NRP1) is garnering renewed attention as it reinforces intratumoral T_reg cell function amidst inflammation in the TME. Loss of NRP1 on T_reg cells in mouse models restores antitumor immunity without sacrificing peripheral tolerance. Enrichment of NRP1⁺ T_reg cells is observed in patients across multiple malignancies with cancer, both intratumorally and in peripheral sites. Thus, targeting NRP1 may safely undermine intratumoral T_reg cell fitness, permitting enhanced inflammatory responses with existing immunotherapies. Furthermore, NRP1 has been recently found to modulate tumor-specific CD8⁺ T cell responses. Emerging data suggest that NRP1 restricts CD8⁺ T cell reinvigoration in response to checkpoint inhibitors, and more importantly, acts as a barrier to the long-term durability of CD8⁺ T cell-mediated tumor immunosurveillance. These novel and distinct regulatory mechanisms present an exciting therapeutic opportunity. This review will discuss the growing literature on NRP1-mediated immune modulation which provides a strong rationale for categorizing NRP1 as both a key checkpoint in the TME as well as an immunotherapeutic target with promise either alone or in combination with current standard of care therapeutic regimens.

Keywords: immunomodulation; immunotherapy; tumor microenvironment.

Check out our biomarker ELISA kits for cancer research 

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-VEGF – BI-VEGF
-Angiopoietin-2 – BI-ANG2

 

Biomedica quality cytokine kits

My name is Franz. I am part of the Biomedica assay development team. We are located right in the heart of Europe.

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Citations:

 The Biomedica human Angiopoietin-2 ELISA (cat.no. BI-ANG2) is described in Nephrology Dialysis Transplantation, Volume 34, Issue Supplement_1, June 2019, gfz103.SP339:

NEW BIOACTIVE ANGIOPOIETIN-2 ELISA ALLOWS THE QUANTIFICATION OF ALL THREE ISOFORMS OF ANGIOPOIETIN-2 IN CHRONIC KIDNEY DISEASE

Ana-Maria Suciu Andreea, Jacqueline Wallwitz, Berg Gabriela, Maria Laber Anna, Himmler Gottfried

INTRODUCTION: Angiopoietin-2 (Ang-2) is an important regulator of the angiopoietin-1/Tie-2 receptor signaling system on endothelial cells during angiogenesis. Disruption of this signaling leads to the loss of endothelial integrity and renders the endothelium response towards a variety of pro-inflammatory cytokines and growth factors. Thus, Ang-2 might lead to vascular micro-inflammation in patients with CKD (chronic kidney disease). Ang-2 levels increase with CKD stage, are associated with fluid overload and abnormal cardiac structure and predict mortality in patients with CKD stages 4–5. Although Ang-2 levels return toward normal after successful kidney transplantation, Ang-2 remains a putative cardiovascular risk factor in this population.

METHODS: An enzyme-linked immunosorbent assay for the detection of all three angiopoietin-2 isoforms in human serum and plasma was developed. Two high quality antibodies are combined in a sandwich test format: As capturing antibody a recombinant monoclonal antibody is used. A biotin-labeled polyclonal affinity-purified antibody serves for detection of the analyte. High resolution epitope mapping of the antibodies via peptide microarray technology allowed the identification of linear antibody epitopes. Technical performance and accuracy of the assay were assessed according to ICH/EMEA guidelines.

RESULTS: Microarray data illustrate the binding of the polyclonal detection antibody to human angiopoietin-2 spotted on a chip. Altogether, seven linear epitopes located N-terminal/at the center of angiopoietin-2 were detected. Most relevant linear epitopes are epitope e2 in the super clustering region and e6 near the fibrinogen-like domain, displaying a twofold higher fluorescent signal than the remaining epitopes. For the recombinant monoclonal antibody no fluorescence was recorded. This antibody recognizes a structural epitope within the C-terminus of angiopoietin-2, which covers the bioactive receptor-binding site of the protein. We expect to detect all described angiopoietin-2 isoforms, as the receptor-binding site is conserved and the majority of the polyclonal antibody epitopes are present in all isoforms. This assay is in conformance with ICH/EMEA/FDA guidelines. Validation data demonstrate its applicability in nephrological disorders including chronic kidney disease. Here we compare an apparently healthy population to chronic kidney disease samples on haemodialysis and kidney transplant samples.

CONCLUSIONS: This new angiopoietin-2 ELISA enables a quick and accurate quantification of all human angiopoietin-2 isoforms that are bioactive in chronic kidney disease and other nephrological disorders.

The Angiopoietin-2 (ANG2) ELISA assay has been fully validated according to international quality guidelines (ICH, EMEA, FDA) and is in line with the required quality specifications (data are shown in the validation file). Assay performance characteristics as accuracy, dilution linearity and parallelism, precision, detection limit and sensitivity, sample stability, specificity including characterization of the angiopoietin-2 antibodies, epitope-mapping were performed.

The Biomedica human VEGF (Vascular Endothelial Growth Factor) ELISA (cat.no. BI-VEGF) is described in the following publication:

NOVEL VEGF-A ELISA ALLOWS SENSITIVE QUANTIFICATION OF HUMAN TOTAL BIOACTIVE VEGF-A.

Osteologie 2020; 29(01): 72. DOI: 10.1055/s-0039-3402888. G Berg, Andreea Ana-Maria Suciu, E Gadermaier, J Wallwitz, G Himmler

Introduction Vascular endothelial growth factor A (VEGF-A), a prominent member of growth factors that regulate angiogenesis and development of normal vasculature, plays an important role in bone development and remodeling. Studies have shown that ossification requires vascularization a priori and that most VEGF in the bone comes from osteoblastic cells. Upon secretion from osteoblasts, VEGF activates endothelial cell migration/proliferation and vessel permeability. Moreover, it regulates osteoclastic differentiation and migration in bone repair. Thus, VEGF represents a relevant therapeutic target. The sensitive measurement of low amounts of circulating VEGF-A found in control cohorts of apparently healthy individuals proves to be difficult. Hence, there is a need for a high-sensitivity assay that reliably measures low VEGF-A concentrations.

Methods We developed a high-sensitivity sandwich ELISA for the detection of human total bioactive VEGF-A using high quality, well-characterized recombinant monoclonal and polyclonal anti-human VEGF-A antibodies. The linear epitopes of the polyclonal detection antibody were mapped with microarray technology. Analyte stability was determined and in accordance with ICH and EMEA guidelines, assay parameters like specificity, dilution linearity, and spike recovery were assessed.

Results We demonstrate that bioactive human VEGF-A can reliably be measured in plasma preparations. In contrast, serum VEGF levels are clearly increased in some samples. This indicates that serum preparation might have an influence on the VEGF amount measured as VEGF can be released from platelets during sample manipulation. Most importantly, we show that samples of apparently healthy individuals are measurable over background. The assay covers a calibration range between 0 and 2000 pg/ml and assay characteristics as well as analyte stability meet the international standards of acceptance. The recombinant capture antibody recognizes a structural epitope in the conserved receptor binding-site of VEGF-A, and thus, specifically binds to all bioactive isoforms of VEGF-A. The polyclonal detection antibody recognizes linear epitopes in the first 120 amino acids of the VEGF-A molecule.

Discussion Our novel VEGF-A ELISA provides a reliable and accurate tool for the quantitative determination of all biologically active VEGF-A isoforms with high sensitivity.

Keywords Angiogenesis, Bone Formation, Growth Factors, Bone Repair, Vascular Endothelial Growth Factor

Of note: The VEGF ELISA assay has been validated according to the guidelines of ICH, EMEA, and FDA.  The VEGF assay validation data show that they correspond to the international quality specifications. Various experiments including parallelism and dilution linearity as well as accuracy, precision, analysis of the stability of samples, and the  characterization of antibodies utilized in the VEGF ELISA assay e.g. epitope-mapping, were performed.

The Biomedica EZ4U cell proliferation & cytotoxicity assay was highlighted in a study demonstrating that fasting improved the therapeutic response in liver cancer. As certain therapies are severely limited by resistance, the results may drive clinical studies aimed at improving therapy response. The results have been published by Jelena Krstic and colleagues in Science Advances in January 2022: Fasting improves therapeutic response in hepatocellular carcinoma through p53-dependent metabolic synergism.

EZ4U Cell Proliferation & Cytotoxicity Assay Highlights

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√  Widely cited  +195 publications

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Fasting improves therapeutic response in hepatocellular carcinoma through p53-dependent metabolic synergism.

Krstic J, Reinisch I, Schindlmaier K, Galhuber M, Riahi Z, Berger N, Kupper N, Moyschewitz E, Auer M, Michenthaler H, Nössing C, Depaoli MR, Ramadani-Muja J, Usluer S, Stryeck S, Pichler M, Rinner B, Deutsch AJA, Reinisch A, Madl T, Chiozzi RZ, Heck AJR, Huch M, Malli R, Prokesch A. Sci Adv. 2022 Jan 21;8(3):eabh2635. doi: 10.1126/sciadv.abh2635. Epub 2022 Jan 21. PMID: 35061544; PMCID: PMC8782451.

Abstract

Cancer cells voraciously consume nutrients to support their growth, exposing metabolic vulnerabilities that can be therapeutically exploited. Here, we show in hepatocellular carcinoma (HCC) cells, xenografts, and patient-derived organoids that fasting improves sorafenib efficacy and acts synergistically to sensitize sorafenib-resistant HCC. Mechanistically, sorafenib acts noncanonically as an inhibitor of mitochondrial respiration, causing resistant cells to depend on glycolysis for survival. Fasting, through reduction in glucose and impeded AKT/mTOR signaling, prevents this Warburg shift. Regulating glucose transporter and proapoptotic protein expression, p53 is necessary and sufficient for the sorafenib-sensitizing effect of fasting. p53 is also crucial for fasting-mediated improvement of sorafenib efficacy in an orthotopic HCC mouse model. Together, our data suggest fasting and sorafenib as rational combination therapy for HCC with intact p53 signaling. As HCC therapy is currently severely limited by resistance, these results should instigate clinical studies aimed at improving therapy response in advanced-stage HCC.

The experimental set up is outlined in the “Material and Methods” section in the publication:  Cell viability of HepG2 cells was analyzed using EZ4U assay (Biomedica Immunoassays).

How does the EZ4U cell proliferation assay work?

The EZ4U assay kit is a simple non-radioactive two to five-hour test that employs tetrazolium salts. These are non-toxic and are reduced to colored formazan. The assay discriminates between living and dead cells as the reduction process requires functional mitochondria that are inactivated within a few minutes after cell death. The assay procedure is identical to the 3H incorporation assays and does not require any change in the test protocol, thus offering the benefit of utilizing a non-radioactive substance.

Of note is the possibility to re-cultivate the cells after determining the cell number. The EZ4U assay has been cited in over 195 publications. Product code: BI-5000.

Method:

√  Water-soluble tetrazolium compound penetrates cell membrane
√  In mitochondria reduction to intense coloured formazan which is also water-soluble
√  Formazan is secreted into culture medium
√  Measured by ELISA reader (450 nm or 492 nm)

Kit specifications:

√  Substrate, lyophilised 10 vials
√  Activator solution, ready to use 1 x 30 ml
√  Sufficient reagents for 10 x 96 tests microtiterplates
√  Sample type: culture medium
√  Sample size: 200 µl / test
√  Detection limit: depending on cell lines. Examples are shown in the product insert
√  Incubation time: 2h – 5h

The EZ4U assay was also highlighted in a recent study:

Syndecan-4 Is a Key Facilitator of the SARS-CoV-2 Delta Variant’s Superior Transmission.

Hudák A, Veres G, Letoha A, Szilák L, Letoha T. Int J Mol Sci. 2022 Jan 12;23(2):796. PMID: 35054983.

Abstract

Emerging SARS-CoV-2 variants pose threats to vaccination campaigns against COVID-19. Being more transmissible than the original virus, the SARS-CoV-2 B.1.617 lineage, named the Delta variant, swept through the world in 2021. The mutations in the Delta’s spike protein shift the protein towards a net positive electrostatic potential. To understand the key molecular drivers of the Delta infection, we investigate the cellular uptake of the Delta spike protein and Delta spike-bearing SARS-CoV-2 pseudoviruses. Specific in vitro modification of ACE2 and syndecan expression enabled us to demonstrate that syndecan-4, the syndecan isoform abundant in the lung, enhances the transmission of the Delta variant by attaching its mutated spike glycoprotein and facilitating its cellular entry. Compared to the wild-type spike, the Delta one shows a higher affinity towards heparan sulfate proteoglycans than towards ACE2. In addition to attachment to the polyanionic heparan sulfate chains, the Delta spike’s molecular interactions with syndecan-4 also involve syndecan-4’s cell-binding domain that mediates cell-to-cell adhesion. Regardless of the complexity of these interactions, exogenously added heparin blocks Delta’s cellular entry as efficiently as syndecan-4 knockdown. Therefore, a profound understanding of the molecular mechanisms underlying Delta infections enables the development of molecularly targeted yet simple strategies to reduce the Delta variant’s spread.

The experimental set up is outlined in the “Material and Methods” section in the publication:  Cellular viability of 293T and 293T-ACE2 cells was measured with EZ4U assay.  

 

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√  Natriuretic Peptides                           NT-proBNP, NT-proANP (for human and rat samples), NT-proCNP

√  Endothelins                                              Big Endothelin-1

√  Wnt inhibitors                                        Sclerostin, Dickkopf-1 (DKK-1)

√  Bone metabilism                                   Osteoprotegerin (OPG), soluble free RANKL (sRANKL)

√  Nephrology & transplant                  Fibroblast growth factor 23 (FGF23 c-terminal and FGF23 intact),
                                                                              Endostatin, Angiopoietin-2 (human, mouse and rat) , Neuropilin-1,
                                                                              Vanin-1 urine (human, mouse and rat), anti-C4d antibody

 

RELATED CITATIONS

Big Endothelin-1 (BigET-1)

Plasma Big Endothelin-1 Level Predicted 5-Year Major Adverse Cardiovascular Events in Patients With Coronary Artery Ectasia. Cai Z, Wang H, Yuan S, Yin D, Song W, Dou K.Front Cardiovasc Med. 2021 Nov 29;8:768431. doi: 10.3389/fcvm.2021.768431. PMID: 34912865; PMCID: PMC8667227.

Abstract

Intracerebral hemorrhage (ICH) is caused by bleeding within the brain. Very few  circulating biomarkers are known to be associated with the risk of ICH. Fibroblast growth factor 23 (FGF23) is a bone-derived protein hormone associated with mortality in patients with heart failure. A recent nested case–control study showed that FGF23 is associated with risk of intracerebral hemorrhage: Fibroblast growth factor 23 is associated with risk of intracerebral hemorrhage. Svensson EH, Söderholm M. Eur J Neurol. 2022 Jan;29(1):114-120. doi: 10.1111/ene.15060. PMID: 34379844.

Abstract

Background and purpose: Fibroblast growth factor 23 (FGF23) is an osteogenic hormone associated with chronic kidney disease and is an emerging risk factor for several cardiovascular diseases. The association of FGF23 with stroke is unclear. The aim of this study was to investigate the association of FGF23 with incident intracerebral hemorrhage (ICH).

Methods: This was a nested case-control study of 220 ICH cases and 244 age- and sex-matched controls from the population-based Malmö Diet and Cancer Study (n = 28,449). Incident ICH cases were ascertained using national registers and classified by bleeding location. Logistic regression was used to study the association of plasma levels of FGF23 with incident ICH, adjusting for potential ICH risk factors. Subgroup analyses were performed for lobar and non-lobar ICH, fatal ICH, ICH with large volume and ICH with poor functional outcome, respectively.

Results: Higher FGF23 levels at baseline were significantly associated with incident ICH. After multivariable adjustment, the odds ratio for the association with all ICH was 1.84 (95% confidence interval [CI] 1.25-2.71, p = 0.002) per doubling of FGF23 concentration. For lobar and non-lobar ICH, odds ratios were 1.73 (95% CI 1.04-2.87, p = 0.035) and 2.13 (95% CI 1.32-3.45, p = 0.002), respectively. FGF23 was also significantly associated with fatal ICH, ICH with large volume and ICH with poor functional outcome.

Conclusions: Higher FGF23 was associated with incident ICH in this nested case-control study. Further studies are required to explore whether the association is causal.

Biomedica offers ready to use FGF23 (C-terminal) and FGF23 intact ELISA kits                     

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Related publications

Plasma FGF23 and the risk of stroke: the Northern Manhattan Study (NOMAS).

Wright CB, Dong C, Stark M, Silverberg S, Rundek T, Elkind MS, Sacco RL, Mendez A, Wolf M. Neurology 2014. 13;82(19):1700-6. PMID: 24706015.

Abstract

Objective: To examine fibroblast growth factor 23 (FGF23) as a risk factor for incident stroke in a racially/ethnically diverse population-based urban cohort.

Methods: Stroke-free Northern Manhattan Study participants with FGF23 measurements (n = 2,525) were followed for a mean of 12 (±5) years to detect incident strokes. We used Cox proportional hazards models to estimate the association of baseline FGF23 with incident total, ischemic, and hemorrhagic stroke.

Results: Median FGF23 was 57 relative units (RU)/mL (interquartile range = 44-81 RU/mL). Each unit increase of natural log-transformed FGF23 conferred a 40% greater overall stroke risk after adjusting for estimated glomerular filtration rate and sociodemographic and vascular risk factors (hazard ratio = 1.4, 95% confidence interval 1.1-1.6, p = 0.004). Penalized spline analysis revealed a linear association with overall stroke risk at ≥90 RU/mL FGF23, compared with <90 RU/mL (hazard ratio = 1.5, 95% confidence interval = 1.2-2.1, p = 0.004). Greater FGF23 conferred a doubling of intracerebral hemorrhage (ICH) risk but no significant increased risk of ischemic stroke. The associations of elevated FGF23 levels with greater risks of overall stroke and ICH events were independent of phosphate and parathyroid hormone levels and were similar among participants without chronic kidney disease.

Conclusions: Elevated FGF23 was a risk factor for overall stroke and ICH events, in particular in a racially and ethnically diverse urban community, independent of chronic kidney disease.

Fibroblast growth factor 23 and risk of incident stroke in community-living adults.

Panwar B, Jenny NS, Howard VJ, Wadley VG, Muntner P, Kissela BM, Judd SE, Gutiérrez OM. Stroke. 2015. 46(2):322-8. doi: 10.1161/STROKEAHA.114.007489. PMID: 25563643.

Abstract

 Background and purpose: Fibroblast growth factor 23 (FGF23) is a hormone that regulates phosphorus and vitamin D metabolism. Elevated FGF23 concentrations are associated with excess risk of cardiovascular disease. Associations of FGF23 with stroke outcomes are less clear.

 Methods: Using a case-cohort study design, we examined the association of baseline plasma FGF23 concentrations with incident stroke in the Reasons for Geographic and Racial Differences in Stroke (REGARDS) study, a cohort of black and white adults aged ≥45 years. FGF23 was measured in 615 participants who developed incident stroke (cases) and in 936 participants randomly selected from the REGARDS cohort (comparison subcohort).

 Results: In multivariable-adjusted models, higher calcium and phosphorus concentrations, lower estimated glomerular filtration rate and higher urine albumin excretion were independently associated with higher FGF23. There was no statistically significant association of FGF23 with risk of all-cause stroke in Cox models adjusted for demographic factors and established stroke risk factors (hazard ratio comparing fourth with first quartile 1.19; 95% confidence interval, 0.78-1.82). In prespecified models stratified by stroke subtypes, there was a graded association of FGF23 with risk of cardioembolic stroke in fully adjusted models (quartile 1, reference; quartile 2 hazard ratio, 1.48; 95% confidence interval, 0.63-3.47; quartile 3 hazard ratio, 1.99; 95% confidence interval, 0.89-4.44; quartile 4 hazard ratio, 2.52; 95% confidence interval, 1.08-5.91). There were no statistically significant associations of FGF23 with other ischemic stroke subtypes or with hemorrhagic strokes.

 Conclusions: Higher FGF23 concentrations were associated with higher risk of cardioembolic but not with other stroke subtypes in community-dwelling adults. Additional studies should delineate reasons for these findings.

Diabetes, high blood pressure and obesity are risk factors for chronic kidney disease (CKD). Roughly 10% of the population worldwide is affected. As the function of the kidneys decline, bone fracture risk increases. Bone fragility is associated with kidney function. Sclerostin is a key molecule in bone metabolism. A novel antibody, blocking the actions of Sclerostin, has become available for treating severe osteoporosis. Whether Sclerostin inhibition is useful in the clinical setting of CKD is discussed in the following review:

Cardiovascular Safety of Anti-Sclerostin Therapy in Chronic Kidney Disease.

Cejka D. Metabolites. 2021 Nov 10;11(11):770. doi: 10.3390/metabo11110770. PMID: 34822428; PMCID: PMC8624769.

Abstract

The significance of sclerostin for bone and cardiovascular health in patients with chronic kidney disease (CKD) is complex and incompletely understood. Experimental evidence suggests that anti-sclerostin therapy shows diminished efficacy on bone in the setting of CKD. Limited clinical evidence suggests that the osteoanabolic and anti-resorptive activity is attenuated, but hypocalcemia is more prevalent in patients with advanced CKD (eGFR < 30 mL/min) treated with anti-sclerostin (romosozumab) therapy as compared to patients without kidney disease. Furthermore, sclerostin is prominently expressed in uremic arteries. Whether the inhibition of sclerostin has adverse effects on cardiovascular health in CKD is currently unknown. This review summarizes the current understanding of the physiology and pathophysiology of sclerostin in CKD, with a focus on the cardiovascular safety of anti-sclerostin therapy in patients with or without CKD.

Check out the Biomedica Sclerostin ELISA kit

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Related publications

Bone Fragility Fractures in CKD Patients.

Pimentel A, Ureña-Torres P, Bover J, Luis Fernandez-Martín J, Cohen-Solal M. Calcif Tissue Int. 2021 Apr;108(4):539-550. doi: 10.1007/s00223-020-00779-z. Epub 2020 Nov 21. PMID: 33219822; PMCID: PMC8052229.

Abstract

Chronic kidney diseases (CKD) are associated with mineral and bone diseases (MBD), including pain, bone loss, and fractures. Bone fragility related to CKD includes the risk factors observed in osteoporosis in addition to those related to CKD, resulting in a higher risk of mortality related to fractures. Unawareness of such complications led to a poor management of fractures and a lack of preventive approaches. The current guidelines of the Kidney Disease Improving Global Outcomes (KDIGO) recommend the assessment of bone mineral density if results will impact treatment decision. In addition to bone density, circulating biomarkers of mineral, serum bone turnover markers, and imaging techniques are currently available to evaluate the fracture risk. The purpose of this review is to provide an overview of the epidemiology and pathogenesis of CKD-associated bone loss. The contribution of the current tools and other techniques in development are discussed. We here propose a current view of how to better predict bone fragility and the therapeutic options in CKD.

Sclerostin in chronic kidney disease-mineral bone disorder think first before you block it!

Brandenburg VM, Verhulst A, Babler A, D’Haese PC, Evenepoel P, Kaesler N. Nephrol Dial Transplant. 2019 Mar 1;34(3):408-414. doi: 10.1093/ndt/gfy129. PMID: 29846712.

Abstract

Canonical Wnt signalling activity is a major player in physiological and adaptive bone metabolism. Wnt signalling is regulated by soluble inhibitors, with sclerostin being the most widely studied. Sclerostin’s main origin is the osteocyte and its major function is blockade of osteoblast differentiation and function. Therefore, sclerostin is a potent inhibitor of bone formation and mineralization. Consequently, blocking sclerostin via human monoclonal antibodies (such as romosozumab) represents a promising perspective for the treatment of (postmenopausal) osteoporosis. However, sclerostin’s physiology and the effects of sclerostin monoclonal antibody treatment are not limited to the skeleton. Specifically, the potential roles of sclerostin in chronic kidney disease (CKD) and associated pathologies covered by the term chronic kidney disease and mineral bone disorder (CKD-MBD), which also includes accelerated cardiovascular calcification, warrant specific attention. CKD-MBD is a complex disease condition in which sclerostin antibodies may interfere at different levels and influence the multiform interplay of hyperparathyroidism, renal osteodystrophy and vascular calcification, but the clinical sequelae remain obscure. The present review summarizes the potential effects of sclerostin blockade in CKD-MBD. We will address and summarize the urgent research targets that are being identified and that need to be addressed before a valid risk-benefit ratio can be established in the clinical setting of CKD.

Sclerostin: a new biomarker of CKD-MBD.

Figurek A, Rroji M, Spasovski G. Int Urol Nephrol. 2020 Jan;52(1):107-113. doi: 10.1007/s11255-019-02290-3. Epub 2019 Oct 14. PMID: 31612420.

Abstract

The causes of the increased cardiovascular risk associated with kidney diseases partly reside in the chronic kidney disease-mineral bone disorder (CKD-MBD) syndrome. Three cardiovascular risk factors [hyperphosphatemia, vascular calcification, and elevated fibroblast growth factor 23 (FGF23)] levels have been discovered within the CKD-MBD over the last decades. In addition, sclerostin is recently presented as a new bone and vascular disease biomarker. This 22-kDa glycoprotein, secreted mainly by osteocytes, is a soluble inhibitor of the canonical Wnt pathway that has a pivotal role in bone biology and turnover. CKD patients are reported with higher levels of sclerostin, and levels decrease during dialysis. Sclerostin is associated with vascular calcification and CV risk in CKD, although data are still controversial. The question whether serum sclerostin has protective or deleterious role in CKD-MBD pathophysiology, and therefore in cardiovascular risk and overall mortality, is still open and needs to be answered. The standardization of assays and the establishment of a clear cut-off values when sclerostin starts to switch from physiological to pathophysiological role have to be another important step. Further research is needed also to define its relationship with other CKD-MBD biomarkers for future diagnostic and therapeutic strategies.

Serum sclerostin in vascular calcification and clinical outcome in chronic kidney disease.

Zeng C, Guo C, Cai J, Tang C, Dong Z. Diab Vasc Dis Res. 2018 Mar;15(2):99-105. doi: 10.1177/1479164117742316. Epub 2017 Nov 23. PMID: 29168393.

Abstract

Sclerostin, a potent soluble inhibitor of the Wnt signalling pathway, is known to inhibit bone formation by suppressing osteocytes differentiation and function. Patients with chronic kidney disease have high levels of serum sclerostin. Sclerostin has been implicated in the pathogenesis of vascular calcification, which may promote the cardiovascular events of morbidity and mortality in chronic kidney disease patients. However, the role of sclerostin in vascular calcification and clinical prognosis in chronic kidney disease remains elusive. While some studies suggested a positive correlation between serum sclerostin and vascular calcification or clinical outcome, other studies showed no or even negative correlation between them. Small sample size, heterogeneity in enrolled patients, discrepancy in anatomical structure examined and differences in the applied assays may be responsible for the discrepant results. Nonetheless, anti-sclerostin antibodies may be a new therapeutic approach to increase bone mass and strength in chronic kidney disease. This review aims to have a better understanding of the relationship of serum sclerostin with vascular calcification and clinical outcome in chronic kidney disease patients, and propose the application of anti-sclerostin therapy in chronic kidney disease.

Related products

Bioactive Sclerostin ELISA kit – cat.no. BI-20472

Osteoporosis is a disease that weakens bone. When women get older, at the time around menopause, bone loss increases. Poor nutrition, lack of exercise, hormonal changes and other factors influence bone health. Early intervention can delay the development of osteoporosis and novel biomarkers may help to identify people at risk.

Bone Health & Osteoporosis – what women should know

BIOMEDICA offers Bone Biomarker ELISA Kits for clinical research

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Related products

Osteoprotegerin (OPG) ELISA  , DKK-1 ELISA ,  Sclerostin ELISA , soluble RANKL ELISA

IL-6 ELISA  , VEGF ELISA ,  Angiopoietin-2 ELISA

Bone Health & Osteoporosis – what women should know

Related publications

The effect of exercise intensity on bone in postmenopausal women (part 2): A meta-analysis.

Kistler-Fischbacher M, Weeks BK, Beck BR. Bone. 2021 Feb;143:115697. doi: 10.1016/j.bone.2020.115697. Epub 2020 Dec 24. PMID: 33357834.

Abstract

Background: Previous reviews have concluded that exercise has only modest effects on bone mineral density (BMD) in postmenopausal women. Despite the well-recognized strong positive relationship between load magnitude and bone response observed from animal research, the majority of human trials have examined the effects of only low to moderate intensity exercise on bone. We speculated that meta-analysing according to intensity may reveal a more potent exercise effect at higher intensity.

Objectives: To determine the effects of low, moderate and high intensity exercise on BMD at the spine and hip in postmenopausal women.

Methods: Electronic databases and reference lists were searched for RCTs that examined the effect of exercise compared to control on DXA-derived lumbar spine, femoral neck or total hip BMD in healthy postmenopausal women. Interventions were classified as low, moderate or high intensity and pooled based on classification. Mean differences (MD) were calculated using random effects models and a risk of bias analysis was undertaken. To determine the effect of different exercise types (resistance and impact training) on BMD outcomes, subgroup analyses for all intensity categories and outcomes were conducted. Separate meta-analyses were undertaken to examine the influence of adding exercise to a bone medication intervention and to examine exercise effects on fracture risk.

Results: Fifty-three trials, testing 63 interventions (19 low, 40 moderate, 4 high intensity) were included. At the lumbar spine, high intensity exercise yielded greater BMD effects (MD = 0.031 g/cm² 95% CI [0.012, 0.049], p = 0.002) than moderate (MD = 0.012 g/cm² 95% CI [0.008, 0.017], p < 0.001) and low intensity (MD = 0.010 g/cm² 95% CI [0.005, 0.015], p < 0.001). Low and moderate intensity exercise was equally effective at the femoral neck (low: 0.011 g/cm² 95% CI [0.006, 0.016], p < 0.001; moderate: 0.011 g/cm² 95% CI [0.007, 0.015], p < 0.001), but no effect of high-intensity exercise was observed. Moderate intensity exercise increased total hip BMD (0.008 g/cm² 95% CI [0.004, 0.012], p < 0.001), but low intensity did not. There were insufficient data to meta-analyse the effect of high intensity exercise at the total hip. Resistance training, potentially in combination with impact training, appears to be the most effective osteogenic stimulus at the spine and hip. Findings from meta-regression analyses were not informative and no influence of exercise on medication efficacy was observed. Risk of bias was mainly low or unclear due to insufficient information reported.

Conclusion: High intensity exercise is a more effective stimulus for lumbar spine BMD than low or moderate intensity, but not femoral neck BMD, however, the latter finding may be due to lack of power. While data from high-intensity exercise interventions are limited, the current comprehensive meta-analysis demonstrates the same positive relationship between load magnitude and bone response in humans that is observed in animal research. Findings have implications for optimal exercise prescription for osteoporosis in postmenopausal women.

Nutrition in the prevention and control of osteoporosis.

Ortega RM, Jiménez Ortega AI, Martínez García RM, Cuadrado Soto E, Aparicio A, López-Sobaler AM. Nutrición en la prevención y el control de la osteoporosis Nutr Hosp. 2021 Jan 13;37(Spec No2):63-66. Spanish. doi: 10.20960/nh.03360. PMID: 32993301.

Abstract

Objective: although osteoporosis develops in advanced stages of life, it must be prevented and stopped from the pediatric age, acting on modifiable factors, especially diet and lifestyle. The objective of this work is to review the latest evidence on nutritional improvements that can help in the prevention and control of the disease. Methods: bibliographic search related to the topic. Results: it is advisable to avoid energy restrictions, especially in postmenopausal women and particularly if they have osteopenia/osteoporosis since, in relation to these pathologies, excess weight may be preferable, rather than underweight. Protein intake higher than the recommended one is beneficial for the bone, provided that the calcium intake is adequate. Excessive intake of sugar and saturated fat should be avoided, but attempts should be made to achieve the nutritional goals set for ω-3 polyunsaturated fatty acids and fiber. It is important to monitor vitamin D status and calcium intake, which is inadequate in high percentages of individuals, as well as improving the contribution of vitamins K, C and group B, and also magnesium, potassium, iron, zinc, copper, fluorine, manganese, silicon and boron, and avoiding the excessive contribution of phosphorus and sodium. Conclusions: osteoporosis is an underdiagnosed pathology and of increasing prevalence. Due to its high morbidity and mortality, prevention is important and, from a nutritional point of view, it is convenient to bring the diet closer to the theoretical ideal. In general, increasing the consumption of dairy products, fish, vegetables and fruits, as well as reducing the consumption of salt, during childhood and throughout life, seems convenient for the bone improvement of most of the population.

Osteoporosis

Compston JE, McClung MR, Leslie WD. Lancet. 2019 Jan 26;393(10169):364-376. doi: 10.1016/S0140-6736(18)32112-3. PMID: 30696576.

 Abstract

Fractures resulting from osteoporosis become increasingly common in women after age 55 years and men after age 65 years, resulting in substantial bone-associated morbidities, and increased mortality and health-care costs. Research advances have led to a more accurate assessment of fracture risk and have increased the range of therapeutic options available to prevent fractures. Fracture risk algorithms that combine clinical risk factors and bone mineral density are now widely used in clinical practice to target high-risk individuals for treatment. The discovery of key pathways regulating bone resorption and formation has identified new approaches to treatment with distinctive mechanisms of action. Osteoporosis is a chronic condition and long-term, sometimes lifelong, management is required. In individuals at high risk of fracture, the benefit versus risk profile is likely to be favourable for up to 10 years of treatment with bisphosphonates or denosumab. In people at a very high or imminent risk of fracture, therapy with teriparatide or abaloparatide should be considered; however, since treatment duration with these drugs is restricted to 18-24 months, treatment should be continued with an antiresorptive drug. Individuals at high risk of fractures do not receive adequate treatment and strategies to address this treatment gap-eg, widespread implementation of Fracture Liaison Services and improvement of adherence to therapy-are important challenges for the future.

The Utility of Biomarkers in Osteoporosis Management.

Garnero P. Mol Diagn Ther. 2017 Aug;21(4):401-418. doi: 10.1007/s40291-017-0272-1. PMID: 28271451.

Abstract

The measurement of bone turnover markers is useful for the clinical investigation of patients with osteoporosis. Among the available biochemical markers, the measurements of serum procollagen type I N-terminal propeptide (PINP) and the crosslinked C-terminal telopeptide (serum CTX) have been recommended as reference markers of bone formation and bone resorption, respectively. The important sources of preanalytical and analytical variability have been identified for both markers, and precise measurement can now be obtained. Reference interval data for PINP and CTX have been generated across different geographical locations, which allows optimum clinical interpretation. However, conventional protein-based markers have some limitations, including a lack of specificity for bone tissue, and their inability to reflect osteocyte activity or periosteal metabolism. Thus, novel markers such as periostin, sclerostin and, sphingosine 1-phosphate have been developed to address some of these shortcomings. Recent studies suggest that the measurements of circulating microRNAs, a new class of marker, may represent early biological markers in osteoporosis. Bone markers have been shown to be a useful adjunct to bone mineral density for identifying postmenopausal women at high risk for fracture. Because levels of bone markers respond rapidly to both anabolic and anticatabolic drugs, they are very useful for investigating the mechanism of action of new therapies and, potentially, for predicting their efficacy to reduce fracture risk.

Liver transplantation has become a routine treatment for children with end stage liver failure. Antibody-mediated rejection of the transplant can be monitored by C4d.  This recent study utilized the Biomedica “anti-C4d antibody”:

Liver Histopathology in Late Protocol Biopsies after Pediatric Liver Transplantation.

Markiewicz-Kijewska M, Szymańska S, Pyzlak M, Kaliciński P, Teisseyre J, Kowalski A, Jankowska I, Czubkowski P, Ismail H. Children (Basel). 2021 Aug 1;8(8):671. doi: 10.3390/children8080671. PMID: 34438562; PMCID: PMC8392008.

Liver Transplant: antibody-mediated rejection monitored by C4d

Anti-C4d Antibody Features

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Abstract

Liver transplantation has become a routine treatment for children with end stage liver failure. Recently, the long term survival of pediatric patients after liver transplantation has improved, with a life expectancy much longer than that of adult recipients, but also with longer exposition of the graft to various injuries, including immunological, inflammatory and others. Biochemical tests, although important, do not always reflect graft injury. The aim of our study was to analyze the histopathology of the graft in late protocol biopsies and correlate it with the clinical and biochemical status of these patients. We analyzed 61 protocol liver biopsies taken from 61 patients. Biopsies were taken 9.03-17.09 years (mean 12.68, median 11.74 years) after transplantation. Liver specimens were examined particularly for the presence and stage of liver fibrosis, inflammation, steatosis, and acute or chronic cellular and humoral rejection. We did not find any abnormalities in 26 (42.6%) liver specimens. None of the patients had signs of cellular or antibody mediated rejection or chronic rejection. In 23 liver biopsies (37.7%), we found non-specific lymphoid infiltrates. Another problem was fibrosis (equal to or more than three on the Ishak scale)-we found it in 17 patients, including seven liver specimens (11.5%) with severe fibrosis (Ishak 5-6). Conclusions: Various pathomorphological abnormalities were found in more than half of patients with a median 11.74 years post-transplant follow-up. Most of them presented normal laboratory liver tests at the same time, suggesting a slow subclinical process leading to pathomorphological abnormalities. No single factor for the development of these abnormalities was found, but our study supports the need for protocol liver biopsies even in patients with normal/almost normal biochemical liver tests.

Liver Transplant: antibody-mediated rejection monitored by C4d

 Related publications

Immunostaining Patterns of Posttransplant Liver Biopsies Using 2 Anti-C4d Antibodies  

Chen L, Himmelfarb EA, Sun M, Choi EK, Fan L, Lai J, Kim CJ, Xu H, Wang HL. Appl Immunohistochem Mol Morphol. 2020 Feb;28(2):146-153. doi: 10.1097/PAI.0000000000000723. PMID: 32044883.  

Citation Biomedica C4d antibody (purchased through Alpco, US, cat. no. BI-RC4D)

 Abstract

Histopathologic diagnosis of antibody-mediated rejection in posttransplant liver biopsies is challenging. The recently proposed diagnostic criteria by the Banff Working Group on Liver Allograft Pathology require positive C4d immunohistochemical staining to establish the diagnosis. However, the reported C4d staining patterns vary widely in different studies. One potential explanation may be due to different antibody preparations used by different investigators. In this study, posttransplant liver biopsies from 69 patients histopathologically diagnosed with acute cellular rejection, chronic rejection, or recurrent hepatitis C were immunohistochemically stained using 2 polyclonal anti-C4d antibodies. On the basis of the distribution of C4d immunoreactivity, 5 different staining patterns were observed: portal vein and capillary, hepatic artery, portal stroma, central vein, and sinusoids. The frequency, extent, and intensity of positive C4d staining with the 2 antibody preparations differed significantly for portal veins/capillaries and central veins, but not for hepatic arteries and portal stroma. Positive sinusoidal staining was seen in only 1 case. There were no significant differences in the frequency, extent, and intensity of positive C4d staining among the acute cellular rejection, chronic rejection, and recurrent hepatitis C groups with the 2 anti-C4d antibodies. These data show that different anti-C4d antibodies can show different staining patterns, which may lead to different interpretation. Caution is thus needed when selecting C4d antibodies for clinical use to aid in the diagnosis of antibody-mediated rejection.

Antibody-Mediated Rejection After Liver Transplant  

Lee M. Gastroenterol Clin North Am. 2017 Jun;46(2):297-309. doi: 10.1016/j.gtc.2017.01.005. PMID: 28506366.

Abstract

Antibody-mediated rejection (AMR) in liver transplants is a field in its infancy compared with its allograft cohorts of the kidney and lung. Acute AMR is diagnosed based on specific clinical and histopathologic criteria: serum donor specific antibodies, C4d staining, histopathologic findings on liver biopsy, and exclusion of other entities. In contrast, the histologic features of chronic AMR are not as specific and it is a more challenging diagnosis to make. Treatments of acute and chronic AMR include some combination of steroids, immune-modulating agents, intravenous immunoglobulin, plasmapheresis, and proteasome inhibitors.

Routine C4d immunohistochemistry in cardiac allografts: Long-term outcomes

Husain AN, Mirza KM, Fedson SE. J Heart Lung Transplant. 2017 Dec;36(12):1329-1335. doi: 10.1016/j.healun.2017.09.004. Epub 2017 Sep 14. PMID: 28988608.

 C4d immunostaining is an independent predictor of cardiac allograft vasculopathy and death in heart transplant recipients

Luk A, Alba AC, Butany J, Tinckam K, Delgado D, Ross HJ. Transpl Int. 2015 Jul;28(7):857-63. doi: 10.1111/tri.12560. Epub 2015 Mar 27. PMID: 25778989.

 

Major depressive disorder (MDD) is a leading psychiatric illness across the world. Around 30-60% of patients with depression do not respond to available anti-depressive treatments. The proinflammatory cytokine IL-6 has a crucial role in the development of MDD. IL-6 is consistently increased in blood samples of MDD patients. Blood IL-6 level correlates with depression scores. Inhibiting IL-6 could offer a new approach in treating depression.

Check out our sensitive IL-6 ELISA kit for clinical serum and plasma samples.

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Blood IL-6 level correlates with Depression Scores 

Role of Interleukin-6 in Depressive Disorder. Int J Mol Sci. 2020 Mar 22;21(6):2194. doi: 10.3390/ijms21062194. PMID: 32235786; PMCID: PMC7139933.

Ting EY et al., J. Int J Mol Sci. 2020 Mar 22;21(6):2194. doi: 10.3390/ijms21062194. PMID: 32235786. Full text link

Abstract link

Major depressive disorder (MDD), which is a leading psychiatric illness across the world, severely affects quality of life and causes an increased incidence of suicide. Evidence from animal as well as clinical studies have indicated that increased peripheral or central cytokine interleukin-6 (IL-6) levels play an important role in stress reaction and depressive disorder, especially physical disorders comorbid with depression. Increased release of IL-6 in MDD has been found to be a factor associated with MDD prognosis and therapeutic response, and may affect a wide range of depressive symptomatology. However, study results of the IL6 genetic effects in MDD are controversial. Increased IL-6 activity may cause depression through activation of hypothalamic-pituitary-adrenal axis or influence of the neurotransmitter metabolism. The important role of neuroinflammation in MDD pathogenesis has created a new perspective that the combining of blood IL-6 and other depression-related cytokine levels may help to classify MDD biological subtypes, which may allow physicians to identify the optimal treatment for MDD patients. To modulate the IL-6 activity by IL-6-related agents, current antidepressive agents, herb medication, pre-/probiotics or non-pharmacological interventions may hold great promise for the MDD patients with inflammatory features.

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VEGF ELISA – BI-VEGF – Product link

Angiopoietin-2 ELISA – BI-ANG2 – Product link

Neuropilin-1 ELISA – BI-20409 – Product link

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 On inflammatory hypothesis of depression: what is the role of IL-6 in the middle of the chaos?

Roohi E et al., J Neuroinflammation. 2021 Feb 16;18(1):45. doi: 10.1186/s12974-021-02100-7. PMID: 33593388; PMCID: PMC7884972. Abstract link

Abstract

Many patients with major depressive disorder (MDD) are reported to have higher levels of multiple inflammatory cytokines including interleukin 6 (IL-6). Recent studies both pre-clinical and clinical have advocated for the functional role of IL-6 in development of MDD and suggested a great potential for targeting this cytokine to open new avenues in pharmacotherapy of depression. The purpose of the present narrative review was to provide an integrated account of how IL-6 may contribute to development of depression. All peer-reviewed journal articles published before July 2020 for each area discussed were searched by WOS, PubMed, MEDLINE, Scopus, Google Scholar, for original research, review articles, and book chapters. Publications between 1980 and July 2020 were included. Alterations in IL-6 levels, both within the periphery and the brain, most probably contribute to depression symptomatology in numerous ways. As IL-6 acts on multiple differing target tissues throughout the body, dysregulation of this particular cytokine can precipitate a multitude of events relevant to depression and blocking its effects can prevent further escalation of inflammatory responses, and potentially pave the way for opening new avenues in diagnosis, treatment, and prevention of this debilitating disorder.

The IL-6 antagonist tocilizumab is associated with worse depression and related symptoms in the medically ill.

Knight JM et al., Transl Psychiatry. 2021 Jan 18;11(1):58. doi: 10.1038/s41398-020-01164-y. PMID: 33462203; PMCID: PMC7812704. Abstract link

 Abstract

Because medical illness is associated with increased inflammation and an increased risk for treatment-resistant major depressive disorder, anti-cytokine therapy may represent a novel, and especially efficacious, treatment for depression. We hypothesized that blockade of the interleukin (IL)-6 signaling pathway with tocilizumab would decrease depression and related symptomatology in a longitudinal cohort of allogeneic hematopoietic stem cell transplantation (HCT) patients, a medically ill population with a significant inflammation and psychopathology. Patients undergoing allogeneic HCT received either a single dose of tocilizumab one day prior to HCT (n = 25), or HCT alone (n = 62). The primary outcome included depressive symptoms at 28 days post HCT; anxiety, fatigue, sleep, and pain were assessed at pretreatment baseline and days +28, +100, and +180 post HCT as secondary outcomes. Multivariate regression demonstrated that preemptive treatment with tocilizumab was associated with significantly higher depression scores at D28 vs. the comparison group (β = 5.74; 95% CI 0.75, 10.73; P = 0.03). Even after adjustment for baseline depressive symptoms, propensity score, and presence of acute graft-versus-host disease (grades II-IV) and other baseline covariates, the tocilizumab-exposed group continued to have significantly higher depression scores compared to the nonexposed group at D28 (β = 4.73; 95% CI 0.64, 8.81; P = 0.02). Despite evidence that IL-6 antagonism would be beneficial, blockade of the IL-6 receptor with tocilizumab among medically ill patients resulted in significantly more-not less-depressive symptoms.

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RELATED PUBLICATIONS

Clinical biomarker validation. Allinson JL. Bioanalysis. 2018 Jun 1;10(12):957-968. doi: 10.4155/bio-2018-0061. Epub 2018 Jun 20. PMID: 29923754.

Abstract

Pre-analytical processes in medical diagnostics: New regulatory requirements and standards. Dagher G, Becker KF, Bonin S, Foy C, Gelmini S, Kubista M, Kungl P, Oelmueller U, Parkes H, Pinzani P, Riegman P, Schröder U, Stumptner C, Turano P, Sjöback R, Wutte A, Zatloukal K.N Biotechnol. 2019 Sep 25;52:121-125. doi: 10.1016/j.nbt.2019.05.002. Epub 2019 May 15. PMID: 31102798.

Abstract

Biomarkers serve as markers for cancer progression and prognosis and represent promising therapeutic targets. Improving our understanding of how cancers originate, grow and spread may help to reduce the risk and burden of the disease. Cancer biomarker research has identified Neuropilin-1 and Semaphorin 4D  as novel protein markers  and promising  therapeutic targets. These proteins can easily be detected in human serum and plasma by ELISA assay.

Innovative Biomarkers in Oncology

Check out our biomarker kits for cancer research link

or have a look at our compete Oncology Biomarker Brochure

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Neuropilin-1 ELISA – BI-20409 – Product link

Semaphorin 4D ELISA – BI-20405 – Product link

Innovative Biomarkers in Oncology 

Related publications

Neuropilins Controlling Cancer Therapy Responsiveness. Full text link

Napolitano V, Tamagnone L. Int J Mol Sci. 2019 Apr 25;20(8):2049. doi: 10.3390/ijms20082049. PMID: 31027288. 

Abstract

Neuropilins (NRPs) are cell surface glycoproteins, acting as co-receptors for secreted Semaphorins (SEMAs) and for members of the vascular endothelial growth factor (VEGF) family; they have been initially implicated in axon guidance and angiogenesis regulation, and more recently in cancer progression. In addition, NRPs have been shown to control many other fundamental signaling pathways, especially mediated by tyrosine kinase receptors (RTKs) of growth factors, such as HGF (hepatocyte growth factor), PDGF (platelet derived growth factor) and EGF (epidermal growth factor). This enables NRPs to control a range of pivotal mechanisms in the cancer context, from tumor cell proliferation and metastatic dissemination, to tumor angiogenesis and immune escape. Moreover, cancer treatment failures due to resistance to innovative oncogene-targeted drugs is typically associated with the activity of alternative RTK-dependent pathways; and neuropilins’ capacity to control oncogenic signaling cascades supports the hypothesis that they could elicit such mechanisms in cancer cells, in order to escape cytotoxic stress and therapeutic attacks. Intriguingly, several studies have recently assayed the impact of NRPs inhibition in combination with diverse anti-cancer drugs. In this minireview, we will discuss the state-of-art about the relevance of NRPs as potential predictive biomarkers of drug response, and the rationale to target these proteins in combination with other anticancer therapies.

Semaphorins as emerging clinical biomarkers and therapeutic targets in cancer. Full text link

Mastrantonio R, You H, Tamagnone L. Theranostics. 2021 Jan 15;11(7):3262-3277. doi: 10.7150/thno.54023. PMID: 33537086; PMCID: PMC7847692.

 Abstract

Semaphorins are a large family of developmental regulatory signals, characterized by aberrant expression in human cancers. These molecules crucially control cell-cell communication, cell migration, invasion and metastasis, tumor angiogenesis, inflammatory and anti-cancer immune responses. Semaphorins comprise secreted and cell surface-exposed molecules and their receptors are mainly found in the Plexin and Neuropilin families, which are further implicated in a signaling network controlling the tumor microenvironment. Accumulating evidence indicates that semaphorins may be considered as novel clinical biomarkers for cancer, especially for the prediction of patient survival and responsiveness to therapy. Moreover, preclinical experimental studies have demonstrated that targeting semaphorin signaling can interfere with tumor growth and/or metastatic dissemination, suggesting their relevance as novel therapeutic targets in cancer; this has also prompted the development of semaphorin-interfering molecules for application in the clinic. Here we will survey, in diverse human cancers, the current knowledge about the relevance of semaphorin family members, and conceptualize potential lines of future research development in this field.

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DKK-1 (Dickkopf-1) ELISA kit – BI-20413 –  Product link

√  Direct measurement – no sample pre-dilution
√  CE marked – for IVD use in the EU
√  Day test – all reagents included
√  Widely cited +160 references!

 DKK1 promotes migration and invasion of non-small cell lung cancer via β-catenin signaling pathway. Full text link

Zhang J, Zhang X, Zhao X, Jiang M, Gu M, Wang Z, Yue W Tumour Biol. 2017. 39(7):1010428317703820. doi: 10.1177/1010428317703820. PMID: 28677426.

 Dickkopf-1: A Promising Target for Cancer Immunotherapy. Full text link

Chu HY, Chen Z, Wang L, Zhang ZK, Tan X, Liu S, Zhang BT, Lu A, Yu Y, Zhang G. Front Immunol. 2021 May 20;12:658097. doi: 10.3389/fimmu.2021.658097. PMID: 34093545; PMCID: PMC8174842.

 Leucine-rich alpha-2-glycoprotein (LRG) ELISA kit – BI-LRG1 –  Product link

√  DAY Test – results in 3.5 h
√  RELIABLE – rigorously validated according to FDA/ICH/EMA guidelines
√  ALL reagents included – controls and sufficient amounts of buffers
√  Specific recombinant epitope-mapped antibodies

Leucine-rich α-2-glycoprotein promotes TGFβ1-mediated growth suppression in the Lewis lung carcinoma cell lines. Full text link

Oncotarget. Takemoto N, Serada S, Fujimoto M, Honda H, Ohkawara T, Takahashi T, Nomura S, Inohara H, Naka T. 2015. 10;6(13):11009-22. doi: 10.18632/oncotarget.3557. PMID: 25826092; PMCID: PMC4484435.

Detection of leucine-rich alpha-2-glycoprotein 1-containing immunocomplexes in the plasma of lung cancer patients with epitope-specific mAbs. Abstract link.

Lázár J, Kovács A, Tornyi I, Takács L, Kurucz I. Cancer Biomark. 2021 Nov 1. doi: 10.3233/CBM-210164. Epub ahead of print. PMID: 34744074.

Check out the Biomedica Cytokine ELISA kits using specific recombinant epitope-mapped antibodies:

√  Interleukin-6 (IL-6)  ELISA  –  BI-IL6 – product link
√  Vasular Endothelial Growth Factor (VEGF) ELISA – BI-VEGF –  product link
√  Angiopoietin-2 ELISA  – BI-ANG2 – product link

See our complete Product Brochure Biomarkers in Oncology

LRG monitors disease activity in inflammatory bowel disease (IBD)  receiving adalimumab

 Inflammatory bowel disease (IBD), including Crohn’s disease and ulcerative colitis affects millions of individuals worldwide. IBD is caused by chronic inflammation  in the gut. Though biologics such as TNF-α inhibitors (e.g. adalimumab) are highly effective in the treatment of IMD they can lose their efficacy over time. A recent report identifies LRG (leucine-rich alpha-2 glycoprotein), a novel serum biomarker, to be useful in monitoring disease activity in IBD patients. LRG is strongly associated with mucosal healing and it better reflects endoscopic activity during adalimumab treatment than C-reactive protein (CRP) and fecal calprotectin (fCal).  Click here to learn more.

Check out the Biomedica LRG ELISA  

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Monitoring IBD disease activity with serum marker LRG

Leucine-Rich Alpha-2 Glycoprotein May Be Predictive of the Adalimumab Trough Level and Antidrug Antibody Development for Patients with Inflammatory Bowel Disease: A Sub-Analysis of the PLANET Study. Yanai S Digestion. 2021;102(6):929-937. doi: 10.1159/000517339. Epub 2021 Jul 16. PMID: 34350873; PMCID: PMC8619792. Full text link .

Abstract link

Introduction: The aim of this study was to examine whether biomarkers are predictive of the adalimumab (ADA) trough level and antidrug antibody development in patients with Crohn’s disease (CD) and ulcerative colitis (UC). Methods: Using data obtained in a prospective, multicenter, observational study (PLANET), we assessed serial changes in a novel biomarker – leucine-rich alpha-2 glycoprotein (LRG) – during ADA treatment for patients with active CD and UC. We measured serum LRG, C-reactive protein (CRP), and fecal calprotectin (fCAL) at weeks 0, 12, 24, and 52. The ADA trough level and anti-ADA antibody (AAA) were also measured at weeks 12 and 52. Correlations between the ADA trough level, AAA, and biomarkers were examined. Results: In all, 34 patients with CD and 47 patients with UC were enrolled. The ADA trough level at week 12 or at the time of ADA withdrawal was 8.5 ± 3.9 in the AAA-negative group (n = 70) and 2.9 ± 2.7 μg/mL in the AAA-positive group (n = 8) (p < 0.0001). The ADA trough level at week 12 or at the time of ADA withdrawal was associated with pretreatment LRG (p = 0.0437 and r = -0.23). Conclusion: LRG, rather than CRP or fCAL, may be a marker for predicting the trough level of ADA for patients with CD and UC treated with ADA.

Monitoring IBD disease activity with serum marker LRG

Related publications:

Leucine-rich alpha-2 glycoprotein is a potential biomarker to monitor disease activity in inflammatory bowel disease receiving adalimumab: PLANET study. Shinzaki S et al., J Gastroenterol. 2021. 56(6):560-569. doi: 10.1007/s00535-021-01793-0. Epub 2021 May 3. PMID: 33942166; PMCID: PMC8137624.

 

Abstract  link

Background: This multicenter prospective study (UMIN000019958) aimed to evaluate the usefulness of serum leucin-rich alpha-2 glycoprotein (LRG) levels in monitoring disease activity in inflammatory bowel disease (IBD). Methods: Patients with moderate-to-severe IBD initiated on adalimumab therapy were enrolled herein. Serum LRG, C-reactive protein (CRP), and fecal calprotectin (fCal) levels were measured at week 0, 12, 24, and 52. Colonoscopy was performed at week 0, 12, and 52 for ulcerative colitis (UC), and at week 0, 24, and 52 for Crohn’s disease (CD). Endoscopic activity was assessed using the Simple Endoscopic Score for Crohn’s Disease (SES-CD) for CD and the Mayo endoscopic subscore (MES) for UC. Results: A total of 81 patients was enrolled. Serum LRG levels decreased along with improvements in clinical and endoscopic outcomes upon adalimumab treatment (27.4 ± 12.6 μg/ml at week 0, 15.5 ± 7.7 μg/ml at week 12, 15.7 ± 9.6 μg/ml at week 24, and 14.5 ± 6.8 μg/ml at week 52), being correlated with endoscopic activity at each time point (SES-CD: r = 0.391 at week 0, r = 0.563 at week 24, r = 0.697 at week 52; MES: r = 0.534 at week 0, r = 0.429 at week 12, r = 0.335 at week 52). Endoscopic activity better correlated with LRG compared to CRP and fCal on pooled analysis at all time points (SES-CD: LRG: r = 0.636, CRP: r = 0.402, fCal: r = 0.435; MES: LRG: r = 0.568, CRP: 0.389, fCal: r = 0.426). Conclusions: Serum LRG is a useful biomarker of endoscopic activity both in CD and UC during the adalimumab treatment.

Leucine-rich alpha-2 glycoprotein as a marker of mucosal healing in inflammatory bowel disease. Yasutomi E et al., Sci Rep. 2021 May 27;11(1):11086. doi: 10.1038/s41598-021-90441-x. PMID: 34045529; PMCID: PMC8160157. Full text link.

 

Abstract link

Leucine-rich alpha-2 glycoprotein (LRG) may be a novel serum biomarker for patients with inflammatory bowel disease. The association of LRG with the endoscopic activity and predictability of mucosal healing (MH) was determined and compared with those of C-reactive protein (CRP) and fecal markers (fecal immunochemical test [FIT] and fecal calprotectin [Fcal]) in 166 ulcerative colitis (UC) and 56 Crohn’s disease (CD) patients. In UC, LRG was correlated with the endoscopic activity and could predict MH, but the performance was not superior to that of fecal markers (areas under the curve [AUCs] for predicting MH: LRG: 0.61, CRP: 0.59, FIT: 0.75, and Fcal: 0.72). In CD, the performance of LRG was equivalent to that of CRP and Fcal (AUCs for predicting MH: LRG: 0.82, CRP: 0.82, FIT: 0.70, and Fcal: 0.88). LRG was able to discriminate patients with MH from those with endoscopic activity among UC and CD patients with normal CRP levels. LRG was associated with endoscopic activity and could predict MH in both UC and CD patients. It may be particularly useful in CD.

 

Therapeutic monitoring of adalimumab at non-trough levels in patients with inflammatory bowel disease. Kato M et al.,  PLoS One. 2021 Jul 9;16(7):e0254548. doi: 10.1371/journal.pone.0254548. PMID: 34242369; PMCID: PMC8270420. Abstract link.

Paget’s disease of bone is a chronic disease of the skeleton. Healthy bone is remodeled in a lifelong process where mature bone tissue is removed and new bone tissue is formed. In Paget´s disease this process is out of balance which causes bones to grow larger and weaker than normal. Paget´s usually affects just one or a few bones. In some parts of the world it is the second most common bone disorder after osteoporosis. Learn more

 

PAGET´S AWARENESS DAY – 11. January

 

BIOMEDICA – Bone Disease Biomarker ELISA kits for clinical research www.bmgrp.com

Check out our brochure on bone biomarkers link

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Related ELISA kits

Osteoprotegerin (OPG) link ; DKK-1 link ; Sclerostin link

Related publications:

Management of Paget’s disease of bone.

Reid IR. Osteoporos Int. 2020 May;31(5):827-837. doi: 10.1007/s00198-019-05259-1. PMID: 31848640. Full text link

 

Abstract link

Paget’s disease is a progressive focal bone condition which can result in pain, low quality of life, deformity and other complications. Disease progression can be halted with potent bisphosphonates, resulting in improvement in both quality of life and pain, and normalisation of scintigraphy, plain radiographs and bone histology. Zoledronate has transformed the treatment of Paget’s disease, producing sustained remissions in almost all patients. Thus, it is now possible to normalise bone cell activity and prevent disease progression at low cost, with one or two intravenous injections of zoledronate, greatly reducing follow-up costs. Patients with Paget’s disease who are symptomatic or at risk of complications should have the opportunity to reap these therapeutic benefits. Potent bisphosphonates are highly effective in halting disease progression in Paget’s disease, but guidelines disagree about treatment indications. The efficacy, safety and low cost of zoledronate recommend its use in any patient who is symptomatic or judged to be at risk of complications from Paget’s disease.

 

Paget’s Disease of Bone.

Gennari L, Rendina D, Falchetti A, Merlotti D. Calcif Tissue Int. 2019 May;104(5):483-500. doi: 10.1007/s00223-019-00522-3. PMID: 30671590. Full text link

 

Abstract link

Paget’s disease of bone (PDB) is a chronic and focal bone disorder, characterized by increased osteoclast-mediated bone resorption and a subsequent compensatory increase in bone formation, resulting in a disorganized mosaic of woven and lamellar bone at one or more affected skeletal sites. As a result, bone pain, noticeable deformities, arthritis at adjacent joints, and fractures can occur. In a small proportion of cases neoplastic degeneration in osteosarcoma, or, less frequently, giant cell tumor has been also described at PDB sites. While recent epidemiological evidences clearly indicate a decrease in the prevalence and the severity of PDB, over the past 2 decades there have been consistent advances on the genetic mechanisms of disease. It is now clear that PDB is a genetically heterogeneous disorder, with mutations in at least two different genes (SQSTM1, ZNF687) and more common predisposing variants. As a counterpart to the genetic hypothesis, the focal nature of lesions, the decline in prevalence rates, and the incomplete penetrance of the disease among family members suggest that one or more environmental triggers may play a role in the pathophysiology of PDB. The exact nature of these triggers and how they might interact with the genetic factors are less understood, but recent experimental data from mice models suggest the implication of paramixoviral infections. The clinical management of PDB has also evolved considerably, with the development of potent aminobisphosphonates such as zoledronic acid which, given as a single intravenous infusion, now allows a long-term disease remission in the majority of patients.

 

Diagnosis and Management of Paget’s Disease of Bone in Adults: A Clinical Guideline.

Ralston SH, Corral-Gudino L, Cooper C, Francis RM, Fraser WD, Gennari L, Guañabens N, Javaid MK, Layfield R, O’Neill TW, Russell RGG, Stone MD, Simpson K, Wilkinson D, Wills R, Zillikens MC, Tuck SP. J Bone Miner Res. 2019 Apr;34(4):579-604. doi: 10.1002/jbmr.3657. PMID: 30803025. Full text link

 

Abstract link  

An evidence-based clinical guideline for the diagnosis and management of Paget’s disease of bone (PDB) was developed using GRADE methodology, by a Guideline Development Group (GDG) led by the Paget’s Association (UK). A systematic review of diagnostic tests and pharmacological and nonpharmacological treatment options was conducted that sought to address several key questions of clinical relevance. Twelve recommendations and five conditional recommendations were made, but there was insufficient evidence to address eight of the questions posed. The following recommendations were identified as the most important: 1) Radionuclide bone scans, in addition to targeted radiographs, are recommended as a means of fully and accurately defining the extent of metabolically active disease in patients with PDB. 2) Serum total alkaline phosphatase (ALP) is recommended as a first-line biochemical screening test in combination with liver function tests in screening for the presence of metabolically active PDB. 3) Bisphosphonates are recommended for the treatment of bone pain associated with PDB. Zoledronic acid is recommended as the bisphosphonate most likely to give a favorable pain response. 4) Treatment aimed at improving symptoms is recommended over a treat-to-target strategy aimed at normalizing total ALP in PDB. 5) Total hip or knee replacements are recommended for patients with PDB who develop osteoarthritis in whom medical treatment is inadequate. There is insufficient information to recommend one type of surgical approach over another. The guideline was endorsed by the European Calcified Tissues Society, the International Osteoporosis Foundation, the American Society of Bone and Mineral Research, the Bone Research Society (UK), and the British Geriatric Society. The GDG noted that there had been a lack of research on patient-focused clinical outcomes in PDB and identified several areas where further research was needed. 

NT-proBNP associated with mortality in COVID-19 patients with no history of heart failure:  The cardiac biomarker N-terminal pro-B-type natriuretic peptide (NT-proBNP) plays a central role in the diagnosis and management of heart failure (HF). A recent report in patients with COVID-19 and no HF history demonstrates that high admission NT-proBNP is associated with higher mortality and healthcare resources utilization. The authors conclude that preventive strategies may be required for these patients.

Learn more: Admission NT-proBNP and outcomes in patients without history of heart failure hospitalized with COVID-19. Link to full text.

Check out the Biomedica NT-proBNP ELISA (cat.no. SK-1204)

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Abstract:  Admission NT-proBNP and outcomes in patients without history of heart failure hospitalized with COVID-19.

Yoo J, Grewal P, Hotelling J, Papamanoli A, Cao K, Dhaliwal S, Jacob R, Mojahedi A, Bloom ME, Marcos LA, Skopicki HA, Kalogeropoulos AP. ESC Heart Fail. 2021. 5:4278-4287. doi: 10.1002/ehf2.13548. Epub 2021 Aug 4. PMID: 34346182; PMCID: PMC8426942.

Aims: We examined the value of N-terminal pro-B-type natriuretic peptide (NT-proBNP) in patients admitted for coronavirus disease 2019 (COVID-19) without prior history of heart failure (HF) or cardiomyopathy. Methods and results: Retrospective cohort of consecutive adults (N = 679; median age 59 years; 38.7% women; 87.5% White; 7.1% Black; 5.4% Asian; 34.3% Hispanic) admitted with documented COVID-19 in an academic centre in Long Island, NY. Admission NT-proBNP was categorized using the European Society of Cardiology Heart Failure Association age-specific criteria for acute presentations. We examined (i) mortality and the composite of death or mechanical ventilation and (ii) out-of-hospital, intensive care unit (ICU)-free, and ventilator-free days at 28 days. Estimates were adjusted for confounders using a lasso selection process. Using age-specific criteria, 417 patients (61.4%) had low, 141 (20.8%) borderline, and 121 (17.8%) high NT-proBNP. Mortality was 5.8%, 20.6%, and 36.4% for patients with low, borderline, and high NT-proBNP, respectively. In lasso-adjusted models, high NT-proBNP was associated with higher mortality [hazard ratio (HR) 2.15; 95% confidence interval (CI) 1.06-4.39; P = 0.034] and composite endpoint rates (HR 1.66; 95%CI 1.04-2.66; P = 0.035). Patients with high NT-proBNP had 32%, 33%, and 33% fewer out-of-hospital, ICU-free, and ventilator-free days compared with low NT-proBNP counterparts. Results were consistent across age, sex, and race, and regardless of coronary artery disease or hypertension, except for stronger mortality signal with high NT-proBNP in women. Conclusions: In patients with COVID-19 and no HF history, high admission NT-proBNP is associated with higher mortality and healthcare resources utilization. Preventive strategies may be required for these patients.

Related publications:

Characterization of NT-proBNP in a large cohort of COVID-19 patients. Link to full text.

Caro-Codón J, Rey JR, Buño A, Iniesta AM, Rosillo SO, Castrejon-Castrejon S, Rodriguez-Sotelo L, Martinez LA, Marco I, Merino C, Martin-Polo L, Garcia-Veas JM, Martinez-Cossiani M, Gonzalez-Valle L, Herrero A, López-de-Sa E, Merino JL; CARD-COVID Investigators. Eur J Heart Fail. 2021. Mar;23(3):456-464. doi: 10.1002/ejhf.2095. Epub 2021 Feb 1. PMID: 33421281; PMCID: PMC8013330.

 Abstract:

Aims: Extensive research regarding the association of troponin and prognosis in coronavirus disease 2019 (COVID-19) has been performed. However, data regarding natriuretic peptides are scarce. N-terminal pro B-type natriuretic peptide (NT-proBNP) reflects haemodynamic stress and has proven useful for risk stratification in heart failure (HF) and other conditions such as pulmonary embolism and pneumonia. We aimed to adequately characterize NT-proBNP concentrations using a large cohort of patients with COVID-19, and to investigate its association with prognosis. Methods and results: Consecutive patients with confirmed severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection and available NT-proBNP determinations, from March 1st to April 20th, 2020 who completed at least 1-month follow-up or died, were studied. Of 3080 screened patients, a total of 396 (mean age 71.8 ± 14.6 years, 61.1% male) fulfilled all the selection criteria and were finally included, with a median follow-up of 53 (18-62) days. Of those, 192 (48.5%) presented NT-proBNP levels above the recommended cut-off for the identification of HF. However, only 47 fulfilled the clinical criteria for the diagnosis of HF. Patients with higher NT-proBNP during admission experienced more frequent bleeding, arrhythmias and HF decompensations. NT-proBNP was associated with mortality both in the whole study population and after excluding patients with HF. A multivariable Cox model confirmed that NT-proBNP was independently associated with mortality after adjusting for all relevant confounders (hazard ratio 1.28, 95% confidence interval 1.13-1.44, per logarithmic unit). Conclusion: NT-proBNP is frequently elevated in COVID-19. It is strongly and independently associated with mortality after adjusting for relevant confounders, including chronic HF and acute HF. Therefore, its use may improve early prognostic stratification in this condition.

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• NT-proANP ELISA (cat.no. BI-20892) Link
• Neuropilin-1 (cat.no. BI-20409) Link ;  NRP1 IN COVID-19 disease read more

Biomedica´s Periostin ELISA kit has recently been featured in a clinical study for lung cancer: Serum Periostin Predicts Survival in Lung Cancer Patients.

Lung adenocarcinoma is the most common type of lung cancer. When first diagnosed, between 30-50% of patients have bone metastasis with poor survival outcome. The extracellular matrix protein Periostin mediates the spreading of lung cancer. In vivo data suggest that bone metastatic lung cancer cells induce Periostin expression that may enhance cancer aggressivity. A recent report in patients with lung adenocarcinoma demonstrates that serum Periostin levels may be used as a prognostic biomarker to predict survival in lung cancer.

Serum Periostin Predicts Survival in Lung Cancer Patients

Learn more: Serum total periostin is an independent marker of overall survival in bone metastases of lung adenocarcinoma. Link to full text.
Massy E, Rousseau JC, Gueye M, Bonnelye E, Brevet M, Chambard L, Duruisseaux M, Borel O, Roger C, Guelminger R, Pialat JB, Gineyts E, Bouazza L, Millet M, Maury JM, Clézardin P, Girard N, Confavreux CB. J Bone Oncol. 2021. 29:100364. doi: 10.1016/j.jbo.2021.100364. PMID: 34150488; PMCID: PMC8190464.

Check out the Biomedica PERIOSTIN ELISA (cat.no. BI-20433)

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Serum total periostin is an independent marker of overall survival in bone metastases of lung adenocarcinoma

Abstract

More than 35% of lung adenocarcinoma patients have bone metastases at diagnosis and have a poor survival. Periostin, a carboxylated matrix protein, mediates lung cancer cell dissemination by promoting epithelial-mesenchymal transition, and is involved in bone response to mechanical stress and bone formation regulation. This suggests that periostin may be used as a biomarker to predict survival in lung cancer patients. Serum periostin was assessed at diagnosis in a prospective cohort of 133 patients with lung adenocarcinoma of all stages. Patients were divided into localized and bone metastatic groups. Both groups were matched to healthy controls. Survival analysis and Cox proportional hazards models were conducted in the total population and in bone metastatic group. The median serum periostin level was higher in bone metastatic (n = 67; median: 1752 pmol/L) than in the localized group (n = 66; 861 pmol/L; p < 0.0001). Patients with high periostin (>median) had a poorer overall survival in the whole population (33.3 weeks vs. NR; p < 0.0001) and the bone metastatic group (24.4 vs. 66.1 weeks; p < 0.001). In multivariate analysis, patients with high periostin had increased risk of death (HR = 2.09, 95%CI [1.06-4.13]; p = 0.03). This was also found in the bone metastatic group (HR = 3.62, 95%CI [1.74-7.52]; p = 0.0005). Immunohistochemistry on bone metastasis biopsies showed periostin expression in the bone matrix and nuclear and cytoplasmic staining in cancer cells. Serum periostin was an independent survival biomarker in all-stage and in bone metastatic lung adenocarcinoma patients. IHC data suggest that periostin might be induced in cancer cells in bone metastatic niche in addition to bone microenvironment expression.

Related publications

High serum levels of periostin are associated with a poor survival in breast cancer. Link to full text.
Rachner TD, Göbel A, Hoffmann O, Erdmann K, Kasimir-Bauer S, Breining D, Kimmig R, Hofbauer LC, Bittner AK.Breast Cancer Res Treat. 2020 Apr;180(2):515-524. doi: 10.1007/s10549-020-05570-0. Epub 2020 Feb 10.PMID: 32040688.

Abstract
Purpose: Periostin is a secreted extracellular matrix protein, which was originally described in osteoblasts. It supports osteoblastic differentiation and bone formation and has been implicated in the pathogenesis of several human malignancies, including breast cancer. However, little is known about the prognostic value of serum periostin levels in breast cancer. Methods: In this study, we analyzed serum levels of periostin in a cohort of 509 primary, non-metastatic breast cancer patients. Disseminated tumor cell (DTC) status was determined using bone marrow aspirates obtained from the anterior iliac crests. Periostin levels were stratified according to several clinical parameters and Pearson correlation analyses were performed. Kaplan-Meier survival curves were assessed by using the log-rank (Mantel-Cox) test. To identify prognostic factors, multivariate Cox regression analyses were used. Results: Mean serum levels of periostin were 505 ± 179 pmol/l. In older patients (> 60 years), periostin serum levels were significantly increased compared to younger patients (540 ± 184 pmol/l vs. 469 ± 167 pmol/l; p < 0.0001) and age was positively correlated with periostin expression (p < 0.0001). When stratifying the cohort according to periostin serum concentrations, the overall and breast cancer-specific mortality were significantly higher in those patients with high serum periostin (above median) compared to those with low periostin during a mean follow-up of 8.5 years (17.7% vs. 11.4% breast cancer-specific death; p = 0.03; hazard ratio 1.65). Periostin was confirmed to be an independent prognostic marker for breast cancer-specific survival (p = 0.017; hazard ratio 1.79). No significant differences in serum periostin were observed when stratifying the patients according to their DTC status. Conclusions: Our findings emphasize the relevance of periostin in breast cancer and reveal serum periostin as a potential marker for disease prediction, independent on the presence of micrometastases.

Overexpression of periostin predicts poor prognosis in non-small cell lung cancer. Link to full text.
Hong LZ, Wei XW, Chen JF, Shi Y.Oncol. Lett. 2013. 6:1595-1603. doi: 10.3892/ol.2013.1590. Epub 2013 Sep 18.PMID: 24273600.

Abstract
The periostin protein, encoded by the POSTN gene, is a component of the extracellular matrix, which is expressed by fibroblasts and has been observed in a variety of human malignancies. The present study aimed to detect the expression of periostin in the tissues of non-small cell lung cancer (NSCLC) patients and benign lung tumors, and to correlate the results with the clinicopathological data of the subjects, in order to evaluate periostin as a potential prognostic marker. In total, 49 NSCLC patients and 6 benign lung tumors were included in this study. The protein level of periostin was detected in paired normal/paratumor/cancer tissues by a western blot analysis and the mRNA level in paired normal/cancer tissues was detected by quantitative polymerase chain reaction (qPCR). The results were then correlated with established biological and prognostic factors. Immunohistochemistry was used to confirm the location of periostin in the NSCLC tissues. Uni- and multivariate analyses were performed using Cox’s proportional hazards regression model. The protein level of periostin was elevated in the cancer tissue of the NSCLC patients compared with the normal (P=0.017) and paratumor (P=0.000) tissues. The expression level in the male patients was much higher than in the female patients at the protein (P=0.001) and mRNA (P=0.010) levels. The mRNA level in the non-adenocarcinoma (non-ADC) patients was much higher than in the adenocarcinoma (ADC) patients (P=0.029). Periostin was demonstrated higher expression at the protein level in the pseudotumors and tuberculosis patients than in the adjacent (P=0.016) and surrounding tissues (P=0.001). Immunostaining indicated that high levels of periostin were present in the mesenchymal areas, but not in the cancer cells themselves. The patients with tumors exhibiting high-level periostin expression showed a significantly shorter survival time (P=0.036, log-rank test). The 3-year survival rate was 81.5% for patients with low-level periostin expression (periostin-L; n=27) and 45.4% for patients with high-level periostin expression (periostin-H; n=22). Similarly, pathological node (pN) status was a significant prognostic marker in the univariate Cox survival analysis. Notably, periostin-H expression was also identified as an independent prognostic factor by the multivariate analysis (P=0.011). These results showed that the overexpression of periostin predicts a poor prognosis, therefore it may be regarded as a novel molecule in the progression and development of NSCLC. The results provide an additional target for the adjuvant treatment of NSCLC.

Predictive and prognostic value of serum periostin in advanced non-small cell lung cancer patients receiving chemotherapy. Zhang Y, Yuan D, Yao Y, Sun W, Shi Y, Su X. Tumour Biol. 2017. 39(5):1010428317698367. doi: 10.1177/1010428317698367. PMID: 28459197. Full text link.

Mutational profiling of bone metastases from lung adenocarcinoma: results of a prospective study (POUMOS-TEC). Confavreux CB, Girard N, Pialat JB, Bringuier PP, Devouassoux-Shisheboran M, Rousseau JC, Isaac S, Thivolet-Bejui F, Clezardin P, Brevet M. Bonekey Rep. 2014. 3:580. doi: 10.1038/bonekey.2014.75. PMID: 25328676; PMCID: PMC4181073. Full text link.

Diagnostic and prognostic value of serum periostin in patients with non-small cell lung cancer. Xu CH, Wang W, Lin Y, Qian LH, Zhang XW, Wang QB, Yu LK. Oncotarget. 2017. 8(12):18746-18753. doi: 10.18632/oncotarget.13004. PMID: 27816968; PMCID: PMC5386644. Full text link.

Related products:

DKK-1 (Dickkopf-1) ELISA kit – BI-20413 –  Product link

• Direct measurement – no sample pre-dilution
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DKK1 promotes migration and invasion of non-small cell lung cancer via β-catenin signaling pathway. Zhang J, Zhang X, Zhao X, Jiang M, Gu M, Wang Z, Yue W Tumour Biol. 2017. 39(7):1010428317703820. doi: 10.1177/1010428317703820. PMID: 28677426. Full text link

Dickkopf-1: A Promising Target for Cancer Immunotherapy. Chu HY, Chen Z, Wang L, Zhang ZK, Tan X, Liu S, Zhang BT, Lu A, Yu Y, Zhang G. Front Immunol. 2021 May 20;12:658097. doi: 10.3389/fimmu.2021.658097. PMID: 34093545; PMCID: PMC8174842. Full text link

Leucine-rich alpha-2-glycoprotein (LRG) ELISA kit – BI-LRG1 –  Product link

• DAY Test – results in 3.5 h
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Leucine-rich α-2-glycoprotein promotes TGFβ1-mediated growth suppression in the Lewis lung carcinoma cell lines. Oncotarget. Takemoto N, Serada S, Fujimoto M, Honda H, Ohkawara T, Takahashi T, Nomura S, Inohara H, Naka T. 2015. 10;6(13):11009-22. doi: 10.18632/oncotarget.3557. PMID: 25826092; PMCID: PMC4484435. Full text link.

Exosomal Leucine-Rich-Alpha2-Glycoprotein 1 Derived from Non-Small-Cell Lung Cancer Cells Promotes Angiogenesis via TGF-β Signal Pathway. Li Z, Zeng C, Nong Q, Long F, Liu J, Mu Z, Chen B, Wu D, Wu H. Mol Ther Oncolytics. 2019. 7;14:313-322. doi: 10.1016/j.omto.2019.08.001. PMID: 31528707; PMCID: PMC6739429. Full text link.

Detection of leucine-rich alpha-2-glycoprotein 1-containing immunocomplexes in the plasma of lung cancer patients with epitope-specific mAbs. Lázár J, Kovács A, Tornyi I, Takács L, Kurucz I. Cancer Biomark. 2021 Nov 1. doi: 10.3233/CBM-210164. Epub ahead of print. PMID: 34744074. Abstract link.

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Blood and urine biomarkers are tools to detect diseases, discover drugs and monitor patients. Biomarker research has identified Endostatin and Vanin-1 as promising novel markers to detect microvascular tissue injuries and renal tubular damage in drug-induced acute kidney injury, respectively. These and other proteins e.g. FGF23 and Periostin can easily be detected by ELISA. Check out our assay portfolio for clinical and preclinical research – novel biomarkers in clinical nephrology: www.bmgrp.com.

Assay Highlights:
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Related Publications:

ENDOSTATIN  FOR THE DETECTION OF ADVANCED MICROVASCULAR KIDNEY DAMAGE AND THE PROGRESSION OF KIDNEY DISEASE

Plasma endostatin predicts kidney outcomes in patients with type 2 diabetes. Chauhan K et al.,  Kidney Int,  2019; 95(2):439-446. Link. “Plasma endostatin was strongly associated with kidney outcomes in type 2 diabetics with preserved eGFR and improved risk discrimination over traditional predictors.”

The association between endostatin and kidney disease and mortality in patients with type 2 diabetes. Carlsson et al., Diabetes Metab, 2016; 42(5):351-357. Link. “In patients with T2D, circulating endostatin levels can predict the progression of kidney disease and mortality independently of established kidney disease markers.”

Endostatin in chronic kidney disease: Associations with inflammation, vascular abnormalities, cardiovascular events and survival.  Kanbay et al., Eur J Intern Med, 2016; 33:81-87. Link. “Endostatin levels are independently associated with incident CVE in CKD patients.”

Elevated plasma levels of endostatin are associated with chronic kidney disease. Chen et al., Am J Nephrol, 2012; 35(4):335-340. Link. “These data indicate that elevated plasma endostatin is strongly and independently associated with CKD.”

VANIN-1   A MARKER FOR DRUG-INDUCED & SPONTANEOUS ACUTE KIDNEY INJURY AND OBSTRUCTIVE & DIABETIC NEPHROPATHY

Urinary vanin-1 associated with chronic kidney disease in hypertensive patients: A pilot study. Hosohata K et al.,  J Clin Hypertens (Greenwich). 2020 Aug;22(8):1458-1465. Link. “ .. urinary vanin-1 is associated with lower eGFR and higher UPCR and UACR, and might be a potential marker of decreased kidney function in hypertensive patients.”  

A Novel Biomarker for Acute Kidney Injury, Vanin-1, for Obstructive Nephropathy: A Prospective Cohort Pilot Study. Washino et al., Int J Mol Sci, 2019; 20(4).  Link. “Urinary Vanin-1 is a useful biomarker to detect and monitor the clinical course of obstructive nephropathy.”

Urinary Vanin-1 as a Novel Biomarker for Early Detection of Drug-Induced Acute Kidney Injury. Hosohata et al., J Pharm Exp Ther, 2002; 341(3):656–662. Link. “… compared with urinary Kim-1 and NGAL, urinary vanin-1 is an earlier and equally sensitive biomarker for drug-induced AKI.”

Vanin-1: A Potential Biomarker for Nephrotoxicant-Induced Renal Injury. Hosohata et al., Toxicology, 2011; 290(1):82–88.  Link. “These results suggest that vanin-1 is a useful and rapid biomarker for renal tubular injury induced by organic solvents.”

Early Detection of Renal Injury Using Urinary Vanin-1 in Rats with Experimental Colitis. Hosohata et al., J App Tox, 2014; 34(2):184–190. Link. “Compared with Kim-1 and MCP-1, vanin-1 might be an earlier biomarker for the detection of renal injury in rats with experimental colitis.”

Proteomic identification of vanin-1 as a marker of kidney damage in a rat model of type 1 diabetic nephropathy. Fugmann T et al.,  Kidney Int. 2011: ;80(3):272-81. Link

FGF23  FOR RISK PREDICTION IN CHRONIC RENAL INSUFFICIENCY AND TO DETERMINE CARDIOVASCULAR RISK IN CKD

Association of Fibroblast Growth Factor 23 with Atrial Fibrillation in Chronic Kidney Disease, From the Chronic Renal Insufficiency Cohort Study. Mehta et al., JAMA Cardiology, 2016; 1(5):548-556. Link . “Elevated FGF23 is independently associated with prevalent and incident atrial fibrillation in patients with mild to severe CKD.”

Fibroblast growth factor 23 in patients with acute dyspnea: Data from the Akershus Cardiac Examination (ACE) 2 Study.  Lyngbakkena et al., Clin Biochem, 2018; 52:41-47 . Link. “Circulating FGF23 concentrations provide incremental prognostic information to established risk indices in patients with acute dyspnea.”

FGF23 and vitamin D metabolism in chronic kidney disease – mineral bone disorder.  Piec et al., Bone Abstracts, 2016; 5:P469. Link. “cFGF23 is raised in patients with CKD as a compensatory response to hyperphosphatemia or phosphate overload.”

Renal and Extrarenal Effects of Fibroblast Growth Factor 23. Vervloet, Nature Reviews, 2019; Nephrology 1(2):109–120. Link. “.. FGF23 is also a valuable biomarker as it predicts risk of a wide variety of clinical events, in particular heart failure.”

PERIOSTIN   A BIOMARKER FOR SEVERITY, PROGRESSION AND RESPONSE TO THERAPY IN HUMAN KIDNEY DISEASE ASSOCIATED TO HYPERTENSION

Identification of periostin as a critical marker of progression/reversal of hypertensive nephropathy. Guerrot et al., PLoS One, 2012; 7(3):e31974. Link.  “… the results identify Periostin as a previously unrecognized marker associated with hypertensive nephropathy.”

Periostin Induces Kidney Fibrosis after Acute Kidney Injury via the p38 MAPK Pathway. An et al., Am J Physiol Renal Physiol, 2019; 316(3):F426-F437. Link. “Periostin promotes kidney fibrosis via the p38 MAPK pathway following acute kidney injury triggered by a hypoxic or ischemic insult. Periostin ablation may protect against chronic kidney disease progression”.

Periostin as a tissue and urinary biomarker of renal injury in type 2 diabetes mellitus. Satirapoj et al., PLoS One, 2015; 17; 10(4):e0124055. Link.  “Urinary periostin is an associated renal derangement in patients with established diabetic nephropathy and it may be used as an early marker of diabetic renal injury.” 

Urinary Periostin Excretion Predicts Renal Outcome in IgA Nephropathy. Hwang et al., Am J Nephrol, 2016; 44(6):481-492. Link.  “POSTN/Cr value at initial diagnosis correlated with renal fibrosis and predicted the renal outcomes in patients with IgAN. It could be a promising urinary biomarker for renal fibrosis.”

Effects of periostin deficiency on kidney aging and lipid metabolism. An JN Aging (Albany NY). 2021. 13(19):22649-22665. Link.

Periostin in the Kidney. Wallace DP. Adv Exp Med Biol. 2019;1132:99-112. Link.

The research status and prospect of Periostin in chronic kidney disease. Jia YY, Yu Y, Li HJ. Ren Fail. 2020 Nov;42(1):1166-1172. Link.

Polycystic Kidney Disease and Renal Fibrosis. Xue C, Mei CL. Adv Exp Med Biol. 2019;1165:81-100. Link.

Periostin Promotes Cell Proliferation and Macrophage Polarization to Drive Repair after AKI. Kormann R J Am Soc Nephrol. 2020; 31(1):85-100. Link.

Kidney Injury Molecule-1 and Periostin Urinary Excretion and Tissue Expression Levels and Association with Glomerular Disease Outcomes. Wu Q Glomerular Dis. 2021 Jun;1(2):45-59. doi: 10.1159/000513166. Epub 2021. Link.

SCLEROSTIN  FOR THE DIAGNOSIS OF HIGH BONE TURNOVER IN CKD AND THE PREDICTION OF CORONARY ARTERY CALCIFICATION

Circulating levels of sclerostin but not DKK1 associate with laboratory parameters of CKD-MBD.  Behets et al., PLOS ONE, 2017; 12(5). Link.  “Sclerostin, as opposed to DKK1, may qualify as a biomarker of CKD-MBD, particularly in dialysis patients.”

Sclerostin serum levels correlate positively with bone mineral density and microarchitecture in haemodialysis patients. Cejka et al., Nephrol Dial Transplant, 2012; 27:226-230.  Link. “Dialysis patients had significantly higher Sclerostin levels than controls.”

Serum Sclerostin and adverse outcomes in nondialyzed chronic kidney disease patients.  Kanbay et al., J Clin Endocrinol, 2014; 99(10):E1854–E1861. Link. “Serum sclerostin values are associated, even after multiple adjustments, with fatal and nonfatal cardiovascular events in a nondialyzed CKD population.”

Relationship between plasma levels of sclerostin, calcium–phosphate disturbances, established markers of bone turnover, and inflammation in haemodialysis patients. Pietrzyk et al., Int Urol Nephrol, 2019; 51(3):519-526. Link. “Increased circulating sclerostin levels seem to reflect slower bone turnover in HD patients. Low levels of sclerostin are associated with vitamin D deficiency and good phosphates alignment.”

Sclerostin in chronic kidney disease-mineral bone disorder think first before you block it! Brandenburg VM et al.,  Nephrol Dial Transplant,  2019; ;34(3):408-414. Link

NT-proBNP saliva analysis for heart failure assessment. Monitoring Heart Failure – SALIVA ANALYSIS of Cardiac Marker NT-proBNP

The cardiac marker NT-pro-brain natriuretic peptide (NT-proBNP) is considered as the gold standard biomarker for the diagnosis and monitoring of heart failure (HF). It is tested in blood samples by immunochemical methods. Saliva analysis has gained importance as an alternative non-invasive and efficient tool for the diagnosis of several diseases.  A recent report investigated the use of the Biomedica ELISA assay for the determination of NT-proBNP in saliva samples collected from HF patients.  First results highlight the potential role of saliva analysis for HF assessment through NT-proBNP monitoring.

 

Learn more:  Determination and stability of N-terminal pro-brain natriuretic peptide in saliva samples for monitoring heart failure.   Link to full text.  

Bellagambi FG, Petersen C, Salvo P, Ghimenti S, Franzini M, Biagini D, Hangouët M, Trivella MG, Di Francesco F, Paolicchi A, Errachid A, Fuoco R, Lomonaco T. Sci Rep. 2021. 22;11(1):13088. doi: 10.1038/s41598-021-92488-2. PMID: 34158583.

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 Abstract

Heart failure (HF) is the main cause of mortality worldwide, particularly in the elderly. N-terminal pro-brain natriuretic peptide (NT-proBNP) is the gold standard biomarker for HF diagnosis and therapy monitoring. It is determined in blood samples by the immunochemical methods generally adopted by most laboratories. Saliva analysis is a powerful tool for clinical applications, mainly due to its non-invasive and less risky sampling. This study describes a validated analytical procedure for NT-proBNP determination in saliva samples using a commercial Enzyme-Linked Immuno-Sorbent Assay. Linearity, matrix effect, sensitivity, recovery and assay-precision were evaluated. The analytical approach showed a linear behaviour of the signal throughout the concentrations tested, with a minimum detectable dose of 1 pg/mL, a satisfactory NT-proBNP recovery (95–110%), and acceptable precision (coefficient of variation ≤ 10%). Short-term (3 weeks) and long-term (5 months) stability of NT-proBNP in saliva samples under the storage conditions most frequently used in clinical laboratories (4, − 20, and − 80 °C) was also investigated and showed that the optimal storage conditions were at − 20 °C for up to 2.5 months. Finally, the method was tested for the determination of NT-proBNP in saliva samples collected from ten hospitalized acute HF patients. Preliminary results indicate a decrease in NT-proBNP in saliva from admission to discharge, thus suggesting that this procedure is an effective saliva-based point-of-care device for HF monitoring.

 

Related publications

NT-ProBNP levels in saliva and its clinical relevance to heart failure. Foo JY, Wan Y, Kostner K, Arivalagan A, Atherton J, Cooper-White J, Dimeski G, Punyadeera C. PLoS One. 2012;7(10):e48452. doi: 10.1371/journal.pone.0048452. PMID: 23119023. Full text link

 Abstract: Current blood based diagnostic assays to detect heart failure (HF) have large intra-individual and inter-individual variations which have made it difficult to determine whether the changes in the analyte levels reflect an actual change in disease activity. Human saliva mirrors the body’s health and well being and ∼20% of proteins that are present in blood are also found in saliva. Saliva has numerous advantages over blood as a diagnostic fluid which allows for a non-invasive, simple, and safe sample collection. The aim of our study was to develop an immunoassay to detect NT-proBNP in saliva and to determine if there is a correlation with blood levels. Methods: Saliva samples were collected from healthy volunteers (n = 40) who had no underlying heart conditions and HF patients (n = 45) at rest. Samples were stored at -80°C until analysis. A customised homogeneous sandwich AlphaLISA((R)) immunoassay was used to quantify NT-proBNP levels in saliva. Results: Our NT-proBNP immunoassay was validated against a commercial Roche assay on plasma samples collected from HF patients (n = 37) and the correlation was r(2) = 0.78 (p<0.01, y = 1.705× +1910.8). The median salivary NT-proBNP levels in the healthy and HF participants were <16 pg/mL and 76.8 pg/mL, respectively. The salivary NT-proBNP immunoassay showed a clinical sensitivity of 82.2% and specificity of 100%, positive predictive value of 100% and negative predictive value of 83.3%, with an overall diagnostic accuracy of 90.6%. Conclusion: We have firstly demonstrated that NT-proBNP can be detected in saliva and that the levels were higher in heart failure patients compared with healthy control subjects. Further studies will be needed to demonstrate the clinical relevance of salivary NT-proBNP in unselected, previously undiagnosed populations.

 

Saliva diagnostics – Current views and directions. Exp Biol Med (Maywood). 2017 Mar;242(5):459-472. doi: 10.1177/1535370216681550. PMID: 27903834; PMCID: PMC5367650. Full text link

 Abstract: In this review, we provide an update on the current and future applications of saliva for diagnostic purposes. There are many advantages of using saliva as a biofluid. Its collection is fast, easy, inexpensive, and non-invasive. In addition, saliva, as a “mirror of the body,” can reflect the physiological and pathological state of the body. Therefore, it serves as a diagnostic and monitoring tool in many fields of science such as medicine, dentistry, and pharmacotherapy. Introduced in 2008, the term “Salivaomics” aimed to highlight the rapid development of knowledge about various “omics” constituents of saliva, including: proteome, transcriptome, micro-RNA, metabolome, and microbiome. In the last few years, researchers have developed new technologies and validated a wide range of salivary biomarkers that will soon make the use of saliva a clinical reality. However, a great need still exists for convenient and accurate point-of-care devices that can serve as a non-invasive diagnostic tool. In addition, there is an urgent need to decipher the scientific rationale and mechanisms that convey systemic diseases to saliva. Another promising technology called liquid biopsy enables detection of circulating tumor cells (CTCs) and fragments of tumor DNA in saliva, thus enabling non-invasive early detection of various cancers. The newly developed technology-electric field-induced release and measurement (EFIRM) provides near perfect detection of actionable mutations in lung cancer patients. These recent advances widened the salivary diagnostic approach from the oral cavity to the whole physiological system, and thus point towards a promising future of salivary diagnostics for personalized individual medicine applications including clinical decisions and post-treatment outcome predictions. Impact statement The purpose of this mini-review is to make an update about the present and future applications of saliva as a diagnostic biofluid in many fields of science such as dentistry, medicine and pharmacotherapy. Using saliva as a fluid for diagnostic purposes would be a huge breakthrough for both patients and healthcare providers since saliva collection is easy, non-invasive and inexpensive. We will go through the current main diagnostic applications of saliva, and provide a highlight on the emerging, newly developing technologies and tools for cancer screening, detection and monitoring.

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FGF23, a novel muscle biomarker detected in the early stages of ALS: Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease characterized by progressive muscle weakness. Currently, there are no biomarkers that can predict prognosis or disease progression.  FGF23 is a protein secreted in the plasma by bone cells.  A recent report identifies skeletal muscle as a potential target of FGF23. In a  large collection of human ALS muscle samples the researchers observed that FGF23 protein is increased up to 50-fold in ALS muscle tissues .This report raises important questions of the role of FGF23 in ALS disease pathology.

 

Learn more: FGF23, a novel muscle biomarker detected in the early stages of ALS. Si Y, Kazamel M, Benatar M, Wuu J, Kwon Y, Kwan T, Jiang N, Kentrup D, Faul C, Alesce L, King PH. Sci Rep. 2021 Jun 8;11(1):12062. doi: 10.1038/s41598-021-91496-6. PMID: 34103575; PMCID: PMC8187665.

 

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FGF23, a novel muscle biomarker detected in the early stages of ALS.

Abstract:

Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease characterized by progressive muscle weakness. Skeletal muscle is a prime source for biomarker discovery since it is one of the earliest sites to manifest disease pathology. From a prior RNA sequencing project, we identified FGF23 as a potential muscle biomarker in ALS. Here, we validate this finding with a large collection of ALS muscle samples and found a 13-fold increase over normal controls. FGF23 was also increased in the SOD1^G93A mouse, beginning at a very early stage and well before the onset of clinical symptoms. FGF23 levels progressively increased through end-stage in the mouse. Immunohistochemistry of ALS muscle showed prominent FGF23 immunoreactivity in the endomysial connective tissue and along the muscle membrane and was significantly higher around grouped atrophic fibers compared to non-atrophic fibers. ELISA of plasma samples from the SOD1^G93A mouse showed an increase in FGF23 at end-stage whereas no increase was detected in a large cohort of ALS patients. In conclusion, FGF23 is a novel muscle biomarker in ALS and joins a molecular signature that emerges in very early preclinical stages. The early appearance of FGF23 and its progressive increase with disease progression offers a new direction for exploring the molecular basis and response to the underlying pathology of ALS.

 

Related publications: 

Improving clinical trial outcomes in amyotrophic lateral sclerosis. Kiernan MC, Vucic S, Talbot K, McDermott CJ, Hardiman O, Shefner JM, Al-Chalabi A, Huynh W, Cudkowicz M, Talman P, Van den Berg LH, Dharmadasa T, Wicks P, Reilly C, Turner MR. Nat Rev Neurol. 2021 Feb;17(2):104-118. doi: 10.1038/s41582-020-00434-z. Epub 2020 Dec 18. PMID: 33340024; PMCID: PMC7747476. Full text link

 

The gut microbiome: a key player in the complexity of amyotrophic lateral sclerosis (ALS). Boddy SL, Giovannelli I, Sassani M, Cooper-Knock J, Snyder MP, Segal E, Elinav E, Barker LA, Shaw PJ, McDermott CJ. BMC Med. 2021 Jan 20;19(1):13. doi: 10.1186/s12916-020-01885-3. PMID: 33468103; PMCID: PMC7816375. Full text link

 

Gene therapy for ALS: A review. Amado DA, Davidson BL. Mol Ther. 2021 Apr 9:S1525-0016(21)00195-7. doi: 10.1016/j.ymthe.2021.04.008. Epub ahead of print. PMID: 33839324. Full text link

 

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Expression of the axon-guidance protein receptor Neuropilin 1 is increased in the spinal cord and decreased in muscle of a mouse model of amyotrophic lateral sclerosis. Körner S et al., Eur J Neurosci. 2019.