• home
• C-Type Natriuretic Peptide (CNP) Induces Bone Growth

Role of C-Type Natriuretic Peptide (CNP) in bone growth and development

CNP is a member of the natriuretic peptide family. It acts as a paracrine hormone which is present in several tissues such as the cardiovascular system, brain and the endothelium. CNP controls various processes including blood pressure, water and mineral balance, and numerous metabolic functions (1). In 1995, CNP was first described as a potent regulator of bone remodeling, underlying its important role in humans (2).

C-Type Natriuretic Peptide Induces Bone Growth

Research has indicated that CNP is predominantly synthesized within the joint cartilage by chondrocytes, which are specialized cells primarily responsible for generating collagen and the extracellular matrix. As a result, the primary and well-established role of CNP is in the growth plate skeletal cartilage, where it plays a crucial role in promoting endochondral growth of the developing skeleton (3, 4).

What is the mechanism of bone growth?

The growth of bones is a complex process involving the formation and remodeling of bone tissue throughout the body. It is regulated by a diverse range of hormones, growth factors, and signaling pathways that interact together to maintain skeletal health and function.  

Bone growth can be broadly classified into two types:

Longitudinal growth primarily takes place in the long bones of the body, such as femur and tibia. It is a process by which bones grow in length, predominantly occurring during childhood and adolescence. Longitudinal growth is facilitated by growth plates, which are cartilaginous areas located at the ends of long bones. As the bones elongate, the cartilage in the growth plates gradually undergoes ossification, transforming into bone tissue.

 Appositional growth, on the other hand, is the process by which bones grow in thickness or diameter. This type of growth occurs throughout life and is regulated by a delicate balance between osteoblasts, cells responsible for bone formation and osteoblasts, cells involved in bone resorption and bone-resorbing cells named osteoclasts.

Numerous factors can influence bone growth, including genetics, nutrition, exercise, and hormonal equilibrium. Hormones such as growth hormone and sex hormones play crucial roles in regulating bone growth and development. One hormone that is of particular significance in this regard is C-type natriuretic peptide (CNP), which is essential for the regulation of bone growth.

Quantification of CNP and NT-proCNP in human blood samples

The measurement of biologically active CNP in human blood samples presents a challenge due to its dynamic role in bone growth, its rapid clearance and low circulating concentrations. However, the inactive, but more stable precursor molecule NT-proCNP, has proven to be a reliable alternative. The amino-terminal propeptide NT-proCNP, is synthesized in the growth plate, has a longer half-life and circulates in equimolar concentrations as CNP. Therefore, NT-proCNP is an excellent candidate for investigating the role of C-Type natriuretic Peptide in bone (1, 3).

A study comparing the measurement of CNP and NT-proCNP in human blood samples revealed that an NT-proCNP assay provides more consistent and reliable results, making it the preferred choice for clinical applications (5).

NT-proCNP can reliably be measured in serum and plasma samples with a conventional ELISA assay 

 BIOMEDICA – NT-proCNP ELISA (cat. no BI-20812) 

• Full validation  – following international quality guidelines
• For human and non-human samples (serum, plasma, cell culture, urine)
• Standard 96-well ELISA assay format- color coded and ready to use reagents
• Developed & manufactured by Biomedica  

 

NT-proCNP serves as a biomarker of linear growth  

Research studies in children has provided evidence showing a strong correlation between the levels of NT-proCNP in blood and the rate of linear growth throughout the entire growth process. In infancy, both males and females exhibit high levels of CNP and NT-proCNP, which subsequently decline in early childhood. However, during puberty, NT-proCNP levels rise again, followed by a decrease during adulthood. The peak levels of NT-proCNP align with the age at which peak height velocity occurs. As a result, the measurement of CNP syntheses through NT-proCNP in blood is highly indicative of linear growth in healthy children at all stages of development, making it an exceptional biomarker for monitoring linear growth (3, 6).

The role of NT-proCNP in bone diseases and genetic disorders

While the involvement of CNP in bone growth during development is well-established, its precise role in the adult skeleton remains uncertain. It is plausible that bone remodeling may contribute to increased plasma levels of CNP in children, given that CNP is expressed in osteoblasts and osteoclasts, which are the cells responsible for bone formation and bone degradation. Moreover, alternations in plasma CNP products have been linked to changes in bone turnover markers in adults (7),

CNP has emerged as a significant player in various types of skeletal dysplasia, a group of over 400 genetic disorders characterized by abnormal bone, joint, and cartilage development. Mutations in genes associated with CNP signaling have been identified in these conditions, including achondroplasia and hypochondroplasia, which are the most prevalent forms of dwarfism (8).

CNP has shown to be a promising therapeutic target for treating growth and skeletal disorders. Only recently, a variant of CNP was approved for treatment of achondroplasia (9). Ongoing clinical trials are currently investigating additional inhibitors of the CNP signaling pathway (10).

Related ELISA assay products

-NT-proANP (cat. no. BI-20492) – assay also suitable for preclinical applications (rat, mouse and other species) – 10µl sample volume

-NT-proBNP (cat. no. SK-1204) – CE-marked, for IVD use in EU

-NT-proBNP rat (cat. no. BI-1204R) – sample values provided

 

Literature

1. Circulating products of C-type natriuretic peptide and links with organ function in health and disease. Prickett TC, A Espiner E. Peptides. 2020. 132:170363.
2. C-type natriuretic peptide increases bone resorption in 1,25-dihydroxyvitamin D3-stimulated mouse bone marrow cultures. Holliday LS, Dean AD, Greenwald JE, Glucks SL. J Biol Chem. 1995. 11;270(32):18983-9.
3. Plasma C-Type Natriuretic Peptide: Emerging Applications in Disorders of Skeletal Growth. Espiner E, Prickett T, Olney R. Horm Res Paediatr. 2018;90(6):345-357.
4. Hormones and osteoporosis update. Effects of natriuretic peptides on endochondral bone growth. Clin Calcium. 2009. 19(7):1003-8. Yasoda A, Nakao K. Japanese. PMID: 19567998.
5. Comparative measurement of CNP and NT-proCNP in human blood samples: a methodological evaluation. Kuehnl A, Pelisek J, Bruckmeier M, Safi W, Eckstein HH. J Negat Results Biomed. 2013. 1;12:7.
6. Amino-terminal propeptide of C-type natriuretic peptide (NTproCNP) predicts height velocity in healthy children. Olney RC, Permuy JW, Prickett TC, Han JC, Espiner EA. Clin Endocrinol (Oxf). 2012. 77(3):416-22.
7. C-type natriuretic peptide forms in adult hyperthyroidism: correlation with thyroid hormones and markers of bone turnover. Schouten BJ, Prickett TC, Hunt PJ, Richards AM, Geffner ME, Olney RC, Espiner EA. Clin Endocrinol (Oxf). 2012. 76(6):790-6.
8. New developments in the management of achondroplasia. Högler W, Ward LM. Wien Med Wochenschr. 2020. 170(5-6):104-111.
9. Expanding horizons of achondroplasia treatment: current options and future developments. Fafilek B, Bosakova M, Krejci P. Osteoarthritis Cartilage. 2022. 30(4):535-544.
10. Emerging drug targets for achondroplasia. Savarirayan R. Expert Opin Ther Targets. 2022. 26(5):389-391.

 

 

 

 

We are happy to introduce our new brochure featuring our Quality Cytokine Assays “BIOMEDICA´s CYTOKINE ELISA kits”  

Product Highlights

• Full validation package
• Ready-to-use prediluted standards
• Color coded reagents & controls included
• Kit calibration with WHO international standard

QUALITY CYTOKINE ELISA KITS FOR AFFORDABLE & EFFICIENT RESEARCH

human Cytokine ELISA Kits  – developed & manufactured by Biomedica

• IL-6 (cat. no. BI-IL6) –  high sensitivity – detectable levels
• VEGF (cat. no. BI-VEGF) –  low sample volume – 10 µl – detecting all known VEGF isoforms
• ANGIOPOIETIN-2 (cat. no. BI-ANG2)    / optimized assay range 

 

Also available mouse/rat Angiopoietin-2 ELISA kit (cat. no. BI-ANG2MR) –  only 5µl sample volume.

 

 

ABOUT CYTOKINES

Cytokines are critical cell signaling molecules that play important roles in the immune response and many other physiological processes.  These proteins are produced by a wide variety of cell types that act on specific target cell receptors to trigger a range of biological responses.

Cytokines are involved in many aspects of the immune response, including inflammation, cell growth and differentiation, and cell death (apoptosis). Cytokines also regulate tissue repair and wound healing. Dysregulation of cytokine production and signaling may contribute to various diseases including autoimmune disorders, allergies, and cancer.

Some common cytokines include interleukins, tumor necrosis factor (TNF), and interferons.

Interleukin-6 (IL-6) is a pleiotropic cytokine that is largely involved in immunomodulatory processes. One of its roles is to respond to infections. As a pro-inflammatory cytokine, IL-6 activates the immune response, recruiting immune cells triggering B and T cell response. IL-6 dysregulation is associated with pathologies involving chronic inflammation, autoimmune diseases, and atherosclerosis. However, IL-6 also has beneficial anti-inflammatory and positive metabolic effects that are released during exercise coupled to a physical adaptation to intense training. In addition to its immune functions, IL-6 is involved in many other physiological processes, including bone remodeling and the nervous system.

Angiopoietin-2 (ANG2) is a protein that plays a critical role in regulating the development and remodeling of blood vessels, a process known as angiogenesis. ANG-2 is primarily produced by endothelial cells, which are cells that line the interior of blood vessels. Under normal physiological conditions the expression of ANG2 is low, which allows the blood vessels to undergo a rapid growth and remodeling. However, in pathological conditions, such as inflammation and cancer, ANG2 expression can be upregulated, leading to excessive angiogenesis and blood vessel destabilization.

Vascular endothelial growth factor (VEGF) is a protein mainly produced by endothelial cells. VEGF stimulates the growth and proliferation of endothelial cells, as well as the migration and differentiation of these cells into new blood vessels. VEGF plays a role in the development of cancer and certain ocular diseases, where it promotes the growth of new blood vessels that feed tumors or damage the retina.

There are several different isoforms of VEGF, each with different properties and functions. For example, VEGF-A is the most well-known cytokine of the VEGF family, and is primarily responsible for promoting angiogenesis. Other isoforms, such as VEGF-B and VEGF-C, have different roles in the body, such as regulating the growth of blood vessels in the heart and lymphatic system, respectively.

Related publications

Harnessing cytokines and chemokines for cancer therapy. Propper DJ, Balkwill FR. Nat Rev Clin Oncol. 2022 Apr;19(4):237-253. doi: 10.1038/s41571-021-00588-9. Epub 2022 Jan 7. PMID: 34997230.

Inflammatory Cytokines and Chemokines as Therapeutic Targets in Heart Failure. Hanna A, Frangogiannis NG. Cardiovasc Drugs Ther. 2020 Dec;34(6):849-863. doi: 10.1007/s10557-020-07071-0. Epub 2020 Sep 9. PMID: 32902739; PMCID: PMC7479403.

Around 7 out of 100,000 people are affected by primary malignant brain tumors. Glioblastoma multiforme (GBM) is the most common type of brain tumor. It grows rapidly and has a low prognosis of approximately 15 months. The  mean age after diagnosis is 63 years (1, 2). Despite significant progress in treatment, there remains a critical need for novel treatment options for GBM. 

Natural Malaria Substance – novel treatment option for brain tumors

Artemisinin, deriving from the plant Artemisia annua is a potent natural substance globally used to treat Malaria. It was discovered in the 1970s from Traditional Chinese Medicines and has since then saved millions of lives (3). Due to its cytotoxic properties it has not only been acknowledged for treating Malaria but also for its potential anticancer properties. With the discovery of Artemisinin and its derivates, researchers have been investigating the mechanisms on how these compounds work e.g. in inhibiting tumor cell proliferation, promoting apoptosis, disrupting caner invasion and metastasis, and preventing angiogenesis (4).

The natural malaria substance Artemisinin  –  a novel treatment option for brain tumors. 

In a recent study using genome-wide screening, researchers have identified mitochondrial function, particularly the biosynthesis of porphyrin, as a crucial pathway responsible for Artemisinin´s cytotoxicity. The potential of this novel treatment option for human brain tumors is currently under investigation (5, 6).

EZ4U / MTT Cell viability assay (cat. no. BI-5000) – BIOMEDICA

 

The Biomedica EZ4U cell viability and cytotoxicity assay was employed in mammalian cell lines in the described study:

A whole-genome scan for Artemisinin cytotoxicity reveals a novel therapy for human brain tumors.

The non-toxic and non-radioactive assay EZ4U / MTT Cell viability assay is based on the conversion of the yellow tetrazolium salt to coloured soluble formazan, which is produced by mitochondrial enzymes. The quantity of formazan produced is in direct proportion to the number of viable, living cells present.

Assay Characteristics

• Method: Reduction of tetrazolium salt to coloured formazan
• Sample type cell culture medium
• Sample size 200 μl / test, 10×96 tests
• Detection limit depending on cell lines Incubation time 2 – 5 h

 

Click here to download our EZ4U “easy for you” – brochure

Literature

1. Brain Tumors. Am J Med. McFaline-Figueroa JR, Lee EQ. 2018 Aug;131(8):874-882. doi: 10.1016/j.amjmed.2017.12.039. Epub 2018 Jan 31. PMID: 29371158.
2. Rising Incidence of Glioblastoma Multiforme in a Well-Defined Population. Grech N, Dalli T, Mizzi S, Meilak L, Calleja N, Zrinzo A Cureus. 2020 May 19;12(5):e8195. doi: 10.7759/cureus.8195. PMID: 32572354; PMCID: PMC7302718.
3. The discovery of artemisinin and the Nobel Prize in Physiology or Medicine. Su XZ, Miller LH. Sci China Life Sci. 2015 Nov;58(11):1175-9. doi: 10.1007/s11427-015-4948-7. PMID: 26481135; PMCID: PMC4966551.
4. Antitumor Research on Artemisinin and Its Bioactive Derivatives. Zhang Y, Xu G, Zhang S, Wang D, Saravana Prabha P, Zuo Z. Nat Prod Bioprospect. 2018 Aug;8(4):303-319. doi: 10.1007/s13659-018-0162-1. Epub 2018 Apr 9. PMID: 29633188; PMCID: PMC6102173.
5. A whole-genome scan for Artemisinin cytotoxicity reveals a novel therapy for human brain tumors. Taubenschmid-Stowers J, Orthofer M, Laemmerer A, Krauditsch C, Rózsová M, Studer C, Lötsch D, Gojo J, Gabler L, Dyczynski M, Efferth T, Hagelkruys A, Widhalm G, Peyrl A, Spiegl-Kreinecker S, Hoepfner D, Bian S, Berger W, Knoblich JA, Elling U, Horn M, Penninger JM. EMBO Mol Med. 2023 Mar 8;15(3):e16959. doi: 10.15252/emmm.202216959. Epub 2023 Feb 6. PMID: 36740985.
6. Achilles heel of glioblastoma. JLP Health and its partners discover combinatorial treatment option for incurable brain tumors. Press Release, JLP Health GmbH, Vienna, Austria, 7 February 2023.

Bone metastases affect over 1.5 million cancer patients globally, making bones a favored metastatic site for solid tumors (1). The presence of skeletal metastases can significantly reduce the quality of life for individuals with advanced cancer, as weakened bones can cause pain, fractures, and increase the risk of mortality (2, 3).

Bones have numerous important functions which include structural support, mobility, hematopoiesis, and mineral storage (4). The health and performance of our bone tissue are overseen by a range of cell types, which include:

• Osteoblasts – cells responsible for creating new bone tissue
• Osteoclasts – cells responsible for breaking down bone tissue
• Osteocytes –  cells within the bone that monitor mechanical loading and regulate the process of bone remodeling 

 

Bone biology- cancer and bone

Cancer and Bone – bone a preferred target site for cancer metastasis

Bone is a favored destination for metastasis in specific types of cancer, whereby cancer cells detach from the primary tumor and disseminate throughout the body. Certain cancers, such as breast, prostate, kidney, lung, ovarian, and thyroid, are especially prone to spread to bone (5).

Cancer and  Bone – the RANK/RANKL/OPG system

The RANK/RANKL/OPG system (receptor activator of the nuclear factor-κB ligand/ Osteoprotegerin) was identified more than 20 years ago and remains a widely researched topic until this day. The interaction between RANK and RANKL is essential for bone metabolism and osteoclast development. OPG acts as a decoy receptor for RANKL. By binding completely to RANKL, OPG obstructs the RANKL-RANK interaction, thereby blocking bone resorption. In addition to its role in regulating bone remodeling, the RANK/RANKL/OPG system is directly implicated in tumor cell development, particularly in the progression of breast and prostate cancer as well as leukemia (6-8).

Inhibiting RANK/RANKL signaling in human cancer

A fully humanized monoclonal antibody has been developed to counteract the effects of RANKL, which has received approval for treating bone loss conditions. The antibody functions by disrupting RANK signaling, preventing osteoclast activation and inhibiting bone resorption (6-8). More recently, inhibiting RANKL has been recognized as a significant checkpoint with the potential to influence anti-tumor response. Consequently, RANKL blockade has a direct impact on bone metastasis. Ongoing clinical and experimental trials are evaluating this emerging therapeutic approach (9).

Studies analyzing serum levels of OPG and RANKL

Serum concentrations of both OPG and soluble RANKL can be accessed via an enzyme-linked immunosorbent assay (ELISA). The predictive value of RANKL/OPG serum levels and disseminated tumor cells in breast cancer patients without metastasis was investigated in 509 patients with primary, nonmetastatic breast cancer. The results demonstrated that RANKL serum levels were significantly elevated in patients who developed bone metastases (10). A different study highlighted the role of OPG as a marker of breast cancer risk in women with a BRCA1 mutation. The authors suggested that OPG levels could be associated with disease risk, potentially serving as a marker for breast cancer risk and improving existing risk prediction models by identifying women at high risk of developing the disease (11).

How are circulating serum levels of Osteoprotegerin and soluble RANKL measured?

Both markers can easily be measured with a conventional ELISA assay.

• Osteoprotegerin (OPG) ELISA (cat. no. BI-20403) – day test, 20µl sample volume/well
• Soluble free RANKL ELISA (cat. no. BI-20462) – highly sensitive and reliable

 

– Widely cited in + 450 publications (citations for OPG, and RANKL)

– Extensively validated ELISA assays following international quality guidelines

High Quality ELISA kits – Developed & Manufactured by Biomedica

Related ELISA kits:  DKK-1, Sclerostin, IL-6, VEGF, Angiopoietin-2

References

1. Biology of Bone Tissue: Structure, Function, and Factors That Influence Bone Cells. Florencio-Silva R, Sasso GR, Sasso-Cerri E, Simões MJ, Cerri PS. Biomed Res Int. 2015;2015:421746. doi: 10.1155/2015/421746. Epub 2015 Jul 13. PMID: 26247020; PMCID: PMC4515490.
2. Understanding the Bone in Cancer Metastasis. Fornetti J, Welm AL, Stewart SA. J Bone Miner Res. 2018 Dec;33(12):2099-2113. doi: 10.1002/jbmr.3618. Epub 2018 Nov 26. PMID: 30476357.
3. Bone as a Preferential Site for Metastasis. JBMR Plus. Sowder ME, Johnson RW. 2019 Jan 15;3(3):e10126. doi: 10.1002/jbm4.10126. PMID: 30918918; PMCID: PMC6419612.
4. Cancer to bone: a fatal attraction. Weilbaecher KN, Guise TA, McCauley LK. Nat Rev Cancer. 2011 Jun;11(6):411-25. doi: 10.1038/nrc3055. Epub 2011 May 19. PMID: 21593787; PMCID: PMC3666847.
5. Bone metastasis: mechanisms, therapies, and biomarkers. Clézardin P, Coleman R, Puppo M, Ottewell P, Bonnelye E, Paycha F, Confavreux CB, Holen I. Physiol Rev. 2021 Jul 1;101(3):797-855. doi: 10.1152/ physrev.00012.2019. Epub 2020 Dec 24. PMID: 33356915.
6. The Roadmap of RANKL/RANK Pathway in Cancer. Casimiro S, Vilhais G, Gomes I, Costa L. Cells. 2021 Aug 4;10(8):1978. doi: 10.3390/cells10081978. PMID: 34440747; PMCID: PMC8393235.
7. RANKL biology: bone metabolism, the immune system, and beyond. Ono T, Hayashi M, Sasaki F, Nakashima T. Inflamm Regen. 2020 Feb 7;40:2. doi: 10.1186/s41232-019-0111-3. PMID: 32047573; PMCID: PMC7006158.
8. RANKL and RANK in Cancer Therapy. Physiology (Bethesda). Onji M, Penninger JM. 2023 May 1;38(3):0. doi: 10.1152/physiol.00020.2022. Epub 2022 Dec 6. PMID: 36473204.
9. The Role of the RANKL/RANK Axis in the Prevention and Treatment of Breast Cancer with Immune Checkpoint Inhibitors and Anti-RANKL. Simatou A, Sarantis P, Koustas E, Papavassiliou AG, Karamouzis MV. Int J Mol Sci. 2020 Oct 14;21(20):7570. doi: 10.3390/ijms21207570. PMID: 33066388; PMCID: PMC7590202.
10. Prognostic Value of RANKL/OPG Serum Levels and Disseminated Tumor Cells in Nonmetastatic Breast Cancer. Rachner TD, Kasimir-Bauer S, Göbel A, Erdmann K, Hoffmann O, Browne A, Wimberger P, Rauner M, Hofbauer LC, Kimmig R, Bittner AK. Clin Cancer Res. 2019 Feb 15;25(4):1369-1378. doi: 10.1158/1078-0432.CCR-18-2482. Epub 2018 Nov 13. PMID: 30425091.
11. Delineating the role of osteoprotegerin as a marker of breast cancer risk among women with a BRCA1 mutation. Park SS, Uzelac A, Kotsopoulos J. Hered Cancer Clin Pract. 2022 Apr 13;20(1):14. doi: 10.1186/s13053-022-00223-3. PMID: 35418083; PMCID: PMC9008947.

 

We offer custom analytical testing services for the measurement of your samples. ANALYTICAL SERVICES – BIOMEDICA –  reliable and flexible:

METHODS:  ELISA  & Luminex Assays, microRNA Analysis, and Glycan Profiling.

Advantages of Using Biomedica Analytical Testing Services:

 

• Flexibility: customized according to your project needs
• Expertise: experienced laboratory staff
• Quality: high quality equipment
• Speed: rapid turn-around time to meet your deadlines
• Results: verified and comprehensive results presented in an analytical report

 

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 Analytical Services – reliable and flexible

We can measure any kind of selected biomarker on your “wish” list by ELISA (enzyme linked immunosorbent assay) for clinical and preclinical samples e.g.

• • Clinical samples (serum, plasma, urine, cell culture supernatants)
  • Pre-clinical samples (e.g. NT-proBNP or NT-proANP measurement in mouse, rat,  rabbit, or other species)  
  • Custom applications – contact us

 

Chronic Kidney Disease (CKD) affects more than 10% of the general population worldwide (1). It is characterized by kidney fibrosis, a progressive process that destroys the kidney (2). An effective treatment to stop the progression of CKD is still lacking. Scientists have now discovered a potential mean to treat CKD by inhibiting Angiopoietin-2 (ANG2), a vascular growth factor (3). Angiopoietin-2 is a potential new treatment target for kidney fibrosis in CKD.

Angiopoietin-2 a new treatment target for kidney fibrosis in CKD

Patients with chronic kidney disease (CKD) have increased plasma levels of Angiopoietin-2 (ANG2).  Studies have shown that ANG2 induces kidney injury and fibrosis in patients with CKD as well as in mouse models of kidney disease (3).  Preventing the progression of renal fibrosis by blocking Angiopoietin-2 could be a new approach in treating chronic kidney disease-associated pathologies.

Based on data of the recent study from Chang et al., on the application of an Angiopoietin-2 inhibitor to mouse models of progressive kidney disease, the scientists found a profound inhibitory effects of ANG2 inhibition on macrophage infiltration and fibrosis (3).  

Future clinical studies will be required to investigate the therapeutic potential on the protective effect of Angiopoietin-2 inhibition on progressive kidney disease. Read more: Angiopoietin-2 inhibition attenuates kidney fibrosis by hindering chemokine C-C motif ligand 2 expression and apoptosis of endothelial cells.

BIOMEDICA developed two assays for the accurate measurement of ANGIOPOIETIN-2 

Mouse / rat ANGIOPOIETIN-2 ELISA  (cat. no. BI-ANG2MR)

• only 5µl sample volume /well
• no predilution of samples required
• kit control included
• sample values provided

 

Human ANGIOPOIETIN-2 ELISA (cat. no. BI-ANG2)

• only 20µl sample volume /well
• the assay is optimized for clinical samples – sample values provided
• ready to use standards and controls  included

 

References

1. Prevalence, outcomes, and cost of chronic kidney disease in a contemporary population of 2·4 million patients from 11 countries: The CaReMe CKD study. Sundström J, Bodegard J, Bollmann A, Vervloet MG, Mark PB, Karasik A, Taveira-Gomes T, Botana M, Birkeland KI, Thuresson M, Jäger L, Sood MM, VanPottelbergh G, Tangri N; CaReMe CKD Investigators. Lancet Reg Health Eur. 2022 Jun 30;20:100438. doi: 10.1016/j.lanepe.2022.100438. PMID: 36090671; PMCID: PMC9459126.

2. Fibrosis in Chronic Kidney Disease: Pathogenesis and Consequences. Panizo S, Martínez-Arias L, Alonso-Montes C, Cannata P, Martín-Carro B, Fernández-Martín JL, Naves-Díaz M, Carrillo-López N, Cannata-Andía JB. Int J Mol Sci. 2021 Jan 2;22(1):408. doi: 10.3390/ijms22010408. PMID: 33401711; PMCID: PMC7795409.

3. Angiopoietin-2 inhibition attenuates kidney fibrosis by hindering chemokine C-C motif ligand 2 expression and apoptosis of endothelial cells. Chang FC, Liu CH, Luo AJ, Tao-Min Huang T, Tsai MH, Chen YJ, Lai CF, Chiang CK, Lin TH, Chiang WC, Chen YM, Chu TS, Lin SL. Kidney Int. 2022 Oct;102(4):780-797. doi: 10.1016/j.kint.2022.06.026. Epub 2022 Aug 4. PMID: 35934136.

Abstract

Plasma levels of angiopoietin-2 are increased in patients with chronic kidney disease (CKD). Moreover, mouse models of progressive kidney disease also demonstrate increased angiopoietin-2 in both plasmas and kidneys. The role of dysregulated angiopoietins in the progression of kidney disease has not been thoroughly investigated. Here, we found in a cohort of 319 patients with CKD that plasma angiopoietin-2 and angiopoietin-2/angiopoietin-1 ratios were positively associated with the development of kidney failure. In mice with progressive kidney disease induced by either ureteral obstruction or ischemia-reperfusion injury, overexpression of human angiopoietin-1 in the kidney tubules not only reduced macrophage infiltration in the initial stage post-injury but also attenuated endothelial cell apoptosis, microvascular rarefaction, and fibrosis in the advanced disease stage. Notably, angiopoietin-1 attenuated chemokine C-C motif ligand 2 (CCL2) expression in the endothelial cells of the fibrosing kidneys, and these protective effects led to attenuation of functional impairment. Mechanistically, angiopoietin-1 reduced CCL2-activated macrophage migration and protected endothelial cells against cell apoptosis induced by angiopoietin-2 and Wnt ligands. Based on this, we applied L1-10, an angiopoietin-2 inhibitor, to the mouse models of progressive kidney disease and found inhibitory effects on macrophage infiltration, microvascular rarefaction, and fibrosis. Thus, we defined the detrimental impact of increased angiopoietin-2 on kidney survival of patients with CKD which appears highlighted by angiopoietin-2 induced endothelial CCL2-activated macrophage infiltration and endothelial cell apoptosis in their kidneys undergoing fibrosis.

Further reading

New therapeutic targets in chronic kidney disease progression and renal fibrosis. Rayego-Mateos S, Valdivielso JM. Expert Opin Ther Targets. 2020 Jul;24(7):655-670. doi: 10.1080/14728222.2020.1762173. Epub 2020 May 18. PMID: 32338087.

 

 

The International Day of Happiness has been initiated by the United Nations to spread awareness of the importance of happiness in the lives of people around the world (1). Happiness and health are closely tied together and affect one another.

Happiness and Health

Numerous studies have found that happiness is linked to a strong immune system and a better general health (2). Reducing risk factors  e.g., tobacco, alcohol, low-nutrition foods and beverages are only some strategies to reduce disease burden (3).

Studying biomarkers may help to understand the mechanism on how the disease originates and could improve diagnosis and disease prognosis (4).

BIOMEDICA specializes on the development of cutting-edge biomarker assays for clinical research.

Biomedica ELISA kits by research area

• Bone Metabolism
• Cardiovascular
• Cell Proliferation
• Immunology
• Infectious Disease
• Metabolic Disease
• Infection
• Nephrology and Transplant
• Neurology
• Oncology
• Oxidative Stress
• Cytokines

High quality ELISA kits- developed and manufactured by Biomedica

References

1. International Day of Happiness – the United Nations  
2. Happiness and Health Annu Rev Public Health. Steptoe A. 2019 Apr 1;40:339-359. doi: 10.1146/annurev-publhealth-040218-044150. Epub 2019 Jan 2. PMID: 30601719. Full text review

Abstract

Research into the relationship between happiness and health is developing rapidly, exploring the possibility that impaired happiness is not only a consequence of ill-health but also a potential contributor to disease risk. Happiness encompasses several constructs, including affective well-being (feelings of joy and pleasure), eudaimonic well-being (sense of meaning and purpose in life), and evaluative well-being (life satisfaction). Happiness is generally associated with reduced mortality in prospective observational studies, albeit with several discrepant results. Confounding and reverse causation are major concerns. Associations with morbidity and disease prognosis have also been identified for a limited range of health conditions. The mechanisms potentially linking happiness with health include lifestyle factors, such as physical activity and dietary choice, and biological processes, involving neuroendocrine, inflammatory, and metabolic pathways. Interventions have yet to demonstrate substantial, sustained improvements in subjective well-being or direct impact on physical health outcomes. Nevertheless, this field shows great potential, with the promise of establishing a favorable effect on population health.

3. Association of Healthy Lifestyle With Years Lived Without Major Chronic Diseases.Nyberg ST, Singh-Manoux A, Pentti J, .. Batty GD, Kivimäki M. JAMA Intern Med. 2020 May 1;180(5):760-768. doi: 10.1001/jamainternmed.  2020.0618. PMID: 32250383; PMCID: PMC7136858.
4. Role of Biomarkers in Health Care. Jain KK. The Handbook of Biomarkers. 2010 Jan 20:115–88. doi: 10.1007/978-1-60761-685-6_5. PMCID: PMC7123449.

 

World Kidney Day – March 9, 2023 – raising awareness of the importance of our kidneys

Around 1 in 10 people suffer from chronic kidney disease (CKD), with more than 800 million individuals being affected worldwide. CKD is a progressive disease in which the kidneys gradually lose their function over time. If detected early, medication and changes in lifestyle may help to prevent or slow down CKD progression.

What are the risk factors for CKD?

Older age (+60 years), diabetes, high blood pressure, heart disease, obesity and some medications are some known risk factors for kidney disease.

How can we keep our kidneys healthy?

Nutrition, exercise, sufficient fluid intake are only some examples on how to keep our kidneys healthy. Valuable information can be found on the following websites :

“World Kidney Day”

“The International Society of Nephrology – ISN”

“International Federation of Kidney Foundations – IFKF”

BIOMEDICA – Biomarker ELISA kits for clinical research in kidney disease

check out our Brochure – Biomarkers in Clinical Nephrology

ROBUST & RELIABLE ASSAYS

• Fully validated according to international quality guidelines
• Widely cited in over 1500 publications

FGF23 (Fibroblast growth factor 23) Vanin-1, Endostatin, Sclerostin, Osteoprotegerin, Angiopoietin-2, IL-6, VEGF

Biomedica Quality ELISA kits

LITERATURE

Plant-based diets for prevention and management of chronic kidney disease. Joshi S, Hashmi S, Shah S, Kalantar-Zadeh K. Curr Opin Nephrol Hypertens. 2020 Jan;29(1):16-21. doi: 10.1097/MNH.0000000000000574. PMID: 31725014. 

Abstract

Purpose of review: Plant-based diets have been used with growing popularity for the treatment of a wide range of lifestyle-related diseases, including diabetes, hypertension, and obesity. With the reinvigoration of the dietary management of chronic kidney disease (CKD) and the use of low protein diets for secondary prevention of CKD to delay or prevent dialysis therapy, there is an increasing interest in the potential role of plant-based diets for these patients.

Recent findings: Recently, a body of evidence related to the role of plant-based diets in preventing CKD has reemerged. Several observational studies have shown that red and processed meat have been associated with increased risk of CKD as well as faster progression in those with preexisting CKD. In several substitution analyses, replacement of one serving of red and/or processed meat has been linked with sizable reductions in CKD risk. Although limited, experimental trials for the treatment of metabolic acidosis in CKD with fruits and vegetables show outcomes comparable to oral bicarbonate. The use of plant-based diets in CKD may have other benefits in the areas of hypertension, weight, hyperphosphatemia, reductions in hyperfiltration, and, possibly, mortality. The risk of potassium overload from plant-based diets appears overstated, mostly opinion-based, and not supported by the evidence. Plant-based diets are generally well tolerated and provide adequate protein intake, including essential amino acids as long as the diet is correctly implemented.

Summary: Plant-based diets should be recommended for both primary and secondary prevention of CKD. Concerns of hyperkalemia and protein inadequacy related to plant-based diets may be outdated and unsupported by the current body of literature. Healthcare providers in general medicine and nephrology can consider plant-based diets as an important tool for prevention and management of CKD.

Exercise and Kidney Disease Prevention: Walk This Way. Seliger S, Weiner DE. Am J Kidney Dis. 2022 Oct;80(4):552-554. doi: 10.1053/j.ajkd.2022.07.001. Epub 2022 Jul 21. PMID: 35872228.

 

Raising awareness, screening and prevention of chronic kidney disease: It takes more than a village. Hsiao LL. Nephrology (Carlton). 2018 Oct;23 Suppl 4:107-111. doi: 10.1111/nep.13459. PMID: 30298651.

Abstract

Chronic kidney disease (CKD) is a major public health problem worldwide. Its prevalence and incidence are increasing, particularly among the ethnic minority populations. Diabetes, hypertension and obesity have been the three major aetiologies for CKD in all developed countries. While diabetes and hypertension remain the major causes of CKD in developing countries, environmental pollution, pesticides, water, analgesic abuse and herbal medications are common causes in these regions. Rapid urbanization and globalization are thought to be the contributing factors to rising prevalence and incidents of CKD. Despite the rising prevalence of CKD, disease awareness remains profoundly low. Worldwide, only 6% of the general population and 10% of the high-risk population are aware of their CKD statuses. Health screenings have been shown to be effective in improving the incidence of ESRD. However, currently there is no effective tool to assess and evaluate the awareness objectively.

Microbiome-metabolome reveals the contribution of gut-kidney axis on kidney disease. Chen YY, Chen DQ, Chen L, Liu JR, Vaziri ND, Guo Y, Zhao YY. J Transl Med. 2019 Jan 3;17(1):5. doi: 10.1186/s12967-018-1756-4. PMID: 30602367; PMCID: PMC6317198.

 

Acute kidney injury (AKI) – also called acute renal failure –  occurs when the kidneys suddenly fail to function due to an injury medication, or an infection. It happens within a few hours or days and is usually accompanied with other medical conditions e.g. blood vessel blockage, diabetes, heart failure, kidney and liver disease, hospitalization (ICU) and other.

FGF23 in Acute Kidney Injury

Fibroblast growth factor 23 (FGF23) is a bone derived hormone that regulates renal phosphate excretion in the kidney. In kidney disease, when the function of the kidney declines, serum phosphate levels rise which subsequently leads to the secretion of FGF-23. High phosphate levels are also common in patients with acute kidney injury (AKI) (1).

FGF23 marker of adverse outcomes in acute kidney injury 

 Fibroblast growth factor 23 (FGF23) levels rise rapidly with acute kidney injury and are associated with the requirement for renal replacement therapy (1-4). FGF-23 levels also have prognostic utility as shown in a large study in over 1500 patients with AKI (3).

Most studies measuring elevated FGF23 levels in AKI patients are performed using FGF23 (C-terminal) assays, which detect intact and C-terminal FGF23. Although the results mirror an increase of FGF23 production they may not necessarily reflect the bioactivity of FGF23 (5).

Quantifying circulating FGF23 levels

Circulating FGF23 levels include the bioactive intact hormone (iFGF23) and the inactive N-terminal and C-terminal fragments. FGF23 can be measured with two commercially available assay types (6). The C-terminal FGF23 assay captures both intact and the c-terminal fragments of FGF23, which result after cleavage. The intact FGF23 assay utilizes antibodies that are located in the N-terminal and in the C-terminal region of the FGF23 hormone, thus capturing only biologically active intact FGF23.

Learn more:  FGF23 – an Overview

BIOMEDICA offers quality FGF23 assays for serum & plasma samples

 

FGF23 (c-terminal) ELISA, cat. no. BI-20702

FGF23 intact ELISA, cat no. BI-20700

 

• developed & manufactured by Biomedica , Austria
• fully validated
• numerous top journal citations

 

Literature

1. Fibroblast Growth Factor 23 and Klotho in AKI. Christov M, Neyra JA, Gupta S, Leaf DE. Semin Nephrol. 2019 Jan;39(1):57-75. doi: 10.1016/j.semnephrol.2018.10.005. PMID: 30606408. 
2. Fibroblast Growth Factor 23 Regulation and Acute Kidney Injury. Zhou W, Simic P, Rhee EP. Nephron. 2022;146(3):239-242. doi: 10.1159/000517734. Epub 2021 Jul 20. PMID: 34284404; PMCID: PMC8770696. 
3. Fibroblast growth factor 23 associates with death in critically ill patients. Leaf DE, Siew ED, Eisenga MF, Singh K, Mc Causland FR, Srivastava A, Ikizler TA, Ware LB, Ginde AA, Kellum JA, Palevsky PM, Wolf M, Waikar SS, Am Clin J Am Soc Nephrol. 2018. 13(4):531–41. 
4. FGF-23 levels in patients with AKI and risk of adverse outcomes. Leaf DE, Wolf M, Waikar SS, Chase H, Christov M, Cremers S, Stern L. Clin J Am Soc Nephrol. 2012. 7(8):1217-23. 
5. Fibroblast Growth Factor 23 and αKlotho in Acute Kidney Injury: Current Status in Diagnostic and Therapeutic Applications. Neyra JA, Hu MC, Moe OW. Nephron. 2020;144(12):665-672. doi: 10.1159/000509856. Epub 2020 Aug 25. PMID: 32841947; PMCID: PMC7708396. 
6. The Measurement and Interpretation of Fibroblast Growth Factor 23 (FGF23) Concentrations. Heijboer AC, Cavalier E. Calcif Tissue Int. 2023. 112(2):258-270.

 

 

NEW – Rat NT-proBNP ELISA assay 

quantitative, reliable and robust for translational research and drug discovery.

Rat NT-proBNP ELISA assay │ BI-1204R

features & benefits

• 10 µl / well,  serum or plasma
• kit control included
• sample values provided

 

NT-proANP ELISA assay │BI-20892

 features & benefits

• 10 µl / well, serum or plasma
• widely cited as cardiovascular safety biomarker in rats

 

“cross-laboratory comparison study detecting serum NT-proANP in rats with the Biomedica NT-proANP ELISA to be technically adequate with acceptable intra- and inter-assay and inter-laboratory precision and accuracy (Vinken et al., 2016).”

All assays are developed and manufactured by Biomedica! 

NT-proBNP & NT-proANP in drug-induced cardiac hypertrophy

The cardiac biomarkers NT-proBNP and NT-proANP are predictive of left ventricular hypertrophy (LVH) and systolic dysfunction in humans (1, 2).

In preclinical settings they have successfully been used to detect cardiotoxicity (3). NT-proBNP and NT-proANP have shown to be useful tools that help to improve the detection of cardiovascular injury early in drug development (4).

REFERENCES

1. Circulating N-terminal pro-atrial natriuretic peptide is an independent predictor of left ventricular hypertrophy in the general population. The Tromsø Study. Schirmer H, Omland T. Eur Heart J. 1999 May;20(10):755-63. doi: 10.1053/euhj.1998.1396. Erratum in: Eur Heart J 1999 Oct;20(19):1439. PMID: 10329067.
2.  An association between N-terminal pro-brain natriuretic protein level and risk of left ventricular hypertrophy in patients without heart failure. Huang L, Huang L, Yu J, Wu X, Zhao J Exp Ther Med. 2020 May;19(5):3259-3266. doi: 10.3892/etm.2020.8598. Epub 2020 Mar 12. PMID: 32266021; PMCID: PMC7132238.
3.  Cardiac Hypertrophy Working Group of the Predictive Safety Testing Consortium. Serum Natriuretic Peptides as Differential Biomarkers Allowing for the Distinction between Physiologic and Pathologic Left Ventricular Hypertrophy. Dunn ME, Manfredi TG, Agostinucci K, Engle SK, Powe J, King NM, Rodriguez LA, Gropp KE, Gallacher M, Vetter FJ, More V, Shimpi P, Serra D, Colton HM Toxicol Pathol. 2017 Feb;45(2):344-352. doi: 10.1177/0192623316634231. Epub 2016 Jul 11. PMID: 27102652.
4.  Integrated approach to early detection of cardiovascular toxicity induced by a ghrelin receptor agonist. Stokes AH, Falls JG, Yoon L, Cariello N, Faiola B, Colton HM, Jordan HL, Berridge BR. Int J Toxicol. 2015 Mar-Apr;34(2):151-61. doi: 10.1177/1091581815573029. Epub 2015 Feb 26. PMID: 25722321.

 

Valvular heart disease is a leading cause of cardiovascular morbidity. Transcatheter aortic valve replacement (TAVR) is a minimally invasive procedure that replaces the aortic valve in patients with aortic stenosis.  Mineral homeostasis, the process that regulates the body’s levels of calcium and phosphorus, plays an important role in the progression of cardiovascular diseases, including calcific aortic valve stenosis. FGF23 is a bone-derived phosphotropic hormone that regulates phosphate and vitamin D metabolism. FGF23 has been shown to be an independent risk factor for CV morbidity and mortality.

FGF23 is a novel risk factor for patients undergoing TAVR

A study by Mirna et al. evaluated the predictive potential of both C-terminal FGF23 (cFGF23) and intact FGF23 (iFGF23) for adverse outcomes in patients undergoing transcatheter aortic valve replacement (TAVR) with regard to renal function. Patients were followed up at 12 months.

The authors demonstrated:

• significant association of high cFGF23 levels with mortality in patients with an estimated glomerular filtration rate (eGFR) ≥45 ml/min/1.73m².
• patients with cFGF23 levels above the cut-off of 6.82 pmol/l had a worse 1-year-mortality
• cFGF23 could be a novel risk factor for patients undergoing TAVR with eGFR ≥45ml/min/1.73m².

 

FGF23 (C-terminal) and FGF23 intact were both measured with an ELISA assay from BIOMEDICA.

 

Assay Highlights

• full validation package
• for serum and plasma
• +50 international references

 

LITERATURE

Serum levels of C-terminal FGF23 (cFGF23) are associated with 1-year-mortality in patients undergoing transcatheter aortic valve replacement (TAVR).

Mirna M, Lauten A, Jirak P, Rezar R, Wernly B, Paar V, Felder TK, Hoppe UC, Motloch LJ, Jung C, Alushi B, Lichtenauer M, Salmhofer H. Eur J Intern Med. 2021. 85:98-107. doi: 10.1016/j.ejim.2020.09.022. Epub 2020 Nov 13. PMID: 33191056.

Abstract

Introduction: Serum levels of FGF23 have been associated with adverse outcomes in cardiovascular diseases in patients with and without impaired renal function. Hence, this study aimed to explore the prognostic relevance of intact FGF23 (iFGF23) and its derivate C-terminal FGF23 (cFGF23) in patients undergoing transcatheter aortic valve replacement (TAVR) with regard to renal function.

Methods: A total of 274 patients undergoing transfemoral TAVR were enrolled in this study. Blood samples were obtained preinterventionally and analyzed for iFGF23 and cFGF23 by means of enzyme linked immunosorbent assay (ELISA). Follow-up was obtained for 12 months.

Results: Serum levels of cFGF23 and iFGF23 both correlated positively with serum creatinine and inversely with estimated glomerular filtration rate (eGFR). Cox regression analysis revealed a significant association of cFGF23 with 1-year-mortality in patients with eGFR ≥45ml/min/1.73m², but not in patients with an eGFR <45ml/min/1.73m². A cut-off was calculated for cFGF23 (6.82 pmol/l) and patients with eGFR ≥45ml/min/1.73m² were retrospectively divided into two groups (above/below cut-off). Patients above the cut-off had a significantly worse 1-year-mortality than patients below the cut-off (33.3% vs. 19.6%; OR 2.05 (95%CI 1.03-4.07), p= 0.038). The association of cFGF23 with 1-year-mortality in patients with eGFR ≥45ml/min/1.73m² remained statistically significant even after correction for possible confounders in a multivariate Cox regression analysis.

Conclusion: cFGF23 could be an individual risk factor for mortality in patients undergoing TAVR with an eGFR ≥45ml/min/1.73m².

 

 

 

The natriuretic peptides NT-proBNP & NT-proANP

Cardiac Biomarkers – kits for clinical & preclinical use

NT-proBNP is released in the heart in response to cardiac wall stretch or pressure overload.   It is considered as the gold standard in heart failure and is useful for both diagnosis and prognosis (1).

NT-proANP is synthesized and stored in atrial myocytes in response to increased intra-atrial  pressure. Elevated circulating levels of NT-proANP are associated with heart failure resulting from increased mechanical stress in the heart during hypertension. In population-based studies, elevated plasma NT-pro-ANP levels predicted the risk of cardiovascular events and death ( 2).

 

Cardiac Biomarkers – kits for clinical & preclinical use

Natriuretic peptides in pre-clinical models as cardiac toxicity biomarkers

Biomarkers are also a valuable tool to explore drug effects in pre-clinical models. The natriuretic peptide NT-proANP has successfully been validated as a cardiovascular safety biomarker in rodents (3). An additional cardiac toxicity biomarker in drug development is NT-proBNP supporting the use of natriuretic peptides to investigate drug-induced cardiac hypertrophy (4, 5).

Biomedica offers a range of top quality ELISA kits for your clinical & preclinical research

 

• NT-proBNP – human (CE-marked in EU) (#SK-1204)
• NT-proBNP – rat (new!)  (#BI-1204R)
• NT-proANP – human, rodent (#BI-20892) citations rat/mouse

 

 

• widely cited in over 400 publications
• kit validation follows international quality guidelines

 

LITERATURE 

1. NT-proBNP: The Gold Standard Biomarker in Heart Failure. McKie PM, Burnett JC Jr. J Am Coll Cardiol. 2016; 6;68(22):2437-2439. 
2. Plasma natriuretic peptide levels and the risk of cardiovascular events and death. Wang TJ, Larson MG, Levy D, Benjamin EJ, Leip EP, Omland T, Wolf PA, Vasan RS. N Engl J Med. 2004; 12;350(7):655-63. 
3. Cross-laboratory analytical validation of the cardiac biomarker NT-proANP in rat. Vinken P, Reagan WJ, Rodriguez LA, Buck WR, Lai-Zhang J, Goeminne N, Barbacci G, Liu R, King NM, Engle SK, Colton H. Pharmacol Toxicol Methods. 2016; 77:58-65.  
4. Evaluation of Cardiac Toxicity Biomarkers in Rats from Different Laboratories. Kim K, Chini N, Fairchild DG, Engle SK, Reagan WJ, Summers SD, Mirsalis JC – Cardiac Hypertrophy Working Group of the Predictive Safety Testing Consortium. Toxicol Pathol. 2016; 44(8):1072-1083. 
5. Serum Natriuretic Peptides as Differential Biomarkers Allowing for the Distinction between Physiologic and Pathologic Left Ventricular Hypertrophy. Dunn ME, Manfredi TG, Agostinucci K, Engle SK, Powe J, King NM, Rodriguez LA, Gropp KE, Gallacher M, Vetter FJ, More V, Shimpi P, Serra D, Colton HM – Cardiac Hypertrophy Working Group of the Predictive Safety Testing Consortium. Toxicol Pathol. 2017; 45(2):344-352.

 

For use in preclinical models – cardiotoxicity biomarkers

Rat NT-proBNP ELISA , cat. no. BI-1204R

NT-proANP ELISA independently validated for rat, cat. no. BI-20892

February is “Heart Month” raising awareness about heart disease. Biomarker research has revolutionized on how heart diseases are diagnosed and treated. Ongoing research on newer exploratory protein biomarkers may shed light into the complex mechanisms that drive the disease process.

Novel Biomarkers for Heart Disease

BIG ENDOTHELIN-1, ENDOSTATIN, FGF23, LRG

 

Big Endothelin-1 (Big ET-1) is a vasoconstrictor peptide and is the precursor of Endothelin-1 (ET-1) the biologically active form. Elevated concentrations provide an independent indicator of heart failure in congestive heart disease (1). Big ET-1 has a longer half-life than ET-1 and circulates in equimolar amounts as ET-1, making it a more reliable biomarker.

Endostatin is a degradation product of collagen XVIII and is an extracellular matrix protein. Endostatin plays an essential role in the pathogenesis of chronic kidney and heart diseases (2). A study in two community-based cohorts of elderly showed an association of high serum endostatin levels with left ventricular dysfunction and an increased heart failure risk (3).  

Fibroblast growth factor-23 (FGF23) is a hormone that regulates phosphate metabolism. FGF23 has been suggested as a novel candidate biomarker of cardiovascular risk. High circulating FGF23 levels have been associated with adverse cardiovascular outcomes such as heart failure and arrhythmias (4).

Leucine-riche alpha-2-glyoprotein (LRG) is a secreted protein that plays an important role in pathogenic ocular neovascularization. LRG is also involved in multiple conditions including cardiovascular disease, diabetes, inflammatory disorders, and cancer. A recent study demonstrated that LRG accurately identify patients with heart failure stronger than BNP, independent of age, sex, and creatinine (5, 6).

NOVEL BIOMARKERS FOR HEART DISEASE

Biomedica offers a range of top quality ELISA kits for your clinical & preclinical research.

-Big Endothelin-1 (#BI-20082H)

-Endostatin (#BI-20742)

-Endostatin mouse/rat (#BI-20742MR)

-FGF23 intact (#BI-20700)

-FGF23 c-terminal (#BI-20702)

-LRG (#BI-LRG)

 

widely cited in over 130 publications kit validation follows international quality guidelines

LITERATURE

1. Plasma concentration of big endothelin-1 and its relation with plasma NT-proBNP and ventricular function in heart failure patients. Rivera M et al., Rev Esp Cardiol. 2005;  58(3):278-84.
2. Endostatin in Renal and Cardiovascular Diseases. Li M et al., Kidney Dis. 2021; 9;7(6):468-481.
3. Circulating endostatin and the incidence of heart failure. Ruge T et al., 2018; 52(5):244-249.
4. Fibroblast Growth Factor 23 and Long-Term Cardiac Function: The Multi-Ethnic Study of Atherosclerosis. Patel RB et al., Cardiovasc Imaging. 2020; 13(11):e011925.
5. Proteomic analysis of coronary sinus serum reveals leucine-rich α2-glycoprotein as a novel biomarker of ventricular dysfunction and heart failure. Watson CJ et al., Circ Heart Fail. 2011; 4(2):188-97.
6. Serum biomarker discovery related to pathogenesis in acute coronary syndrome by proteomic approach  Shin M et al.,Biosci Rep. 2021; 25;41(6):BSR20210344.

Widely cited non-radioactive EZ4U – Cell Proliferation & Cytotoxicity Assay – single step incubation for use on living cells

The Biomedica EZ4U cell proliferation and cytotoxicity assay  was highlighted in a recent study investigating the cytotoxicity of drug combinations tested against different pancreatic cell lines.  Learn more: Cytotoxicity of combinations of the pan-KRAS SOS1 inhibitor BAY-293 against pancreatic cancer cell lines.

Pancreatic ductal adenocarcinomas (PDACs), the most prevalent pancreatic cancer, is highly aggressive with a 5-year overall survival rate of less than 8%.  Late detection, it´s metastatic spread and the resistance to chemotherapy are among some of the factors responsible for poor patient outcome. The PDAC incidence rate is increasing and is particularly related to the general aging of our society. Furthermore, life style habits that include abuse of alcohol and tobacco as well as obesity and type 2 diabetes increases the risk for various types of cancer including PDAC.

EZ4U – Cell Proliferation & Cytotoxicity Assay (cat.no. BI-5000)

-Non-radioactive & non-toxic assay

-Reliable & Sensitive

-Convenient single-step incubation  – for use on living cells

-Widely cited in more than 230 publications

BROCHURE – EZ4U cell proliferation and cytotoxicity assay

 

FURTHER READING

Pancreatic ductal adenocarcinoma: Treatment hurdles, tumor microenvironment and immunotherapy.

World J Gastrointest Oncol. Sarantis P, Koustas E, Papadimitropoulou A, Papavassiliou AG, Karamouzis MV. 2020 Feb 15;12(2):173-181. doi: 10.4251/wjgo.v12.i2.173. PMID: 32104548; PMCID: PMC7031151.

 

Abstract

Pancreatic ductal adenocarcinoma (PDAC) is one of the most lethal diseases, with an average 5-year survival rate of less than 10%. Unfortunately, the majority of patients have unresectable, locally advanced, or metastatic disease at the time of diagnosis. Moreover, traditional treatments such as chemotherapy, surgery, and radiation have not been shown to significantly improve survival. Recently, there has been a swift increase in cancer treatments that incorporate immunotherapy-based strategies to target all the stepwise events required for tumor initiation and progression. The results in melanoma, non-small-cell lung cancer and renal cell carcinoma are very encouraging. Unfortunately, the application of checkpoint inhibitors, including anti-CTLA4, anti-PD-1, and anti-PD-L1 antibodies, in pancreatic cancer has been disappointing. Many studies have revealed that the PDAC microenvironment supports tumor growth, promotes metastasis and consists of a physical barrier to drug delivery. Combination therapies hold great promise for enhancing immune responses to achieve a better therapeutic effect. In this review, we provide an outline of why pancreatic cancer is so lethal and of the treatment hurdles that exist. Particular emphasis is given to the role of the tumor microenvironment, and some of the latest and most promising studies on immunotherapy in PDAC are also presented.

 

Pancreatic ductal adenocarcinoma: biological hallmarks, current status, and future perspectives of combined modality treatment approaches.

Orth M, Metzger P, Gerum S, Mayerle J, Schneider G, Belka C, Schnurr M, Lauber K. Radiat Oncol. 2019 Aug 8;14(1):141. doi: 10.1186/s13014-019-1345-6. PMID: 31395068; PMCID: PMC6688256.

Abstract

Pancreatic ductal adenocarcinoma (PDAC) is a highly devastating disease with poor prognosis and rising incidence. Late detection and a particularly aggressive biology are the major challenges which determine therapeutic failure. In this review, we present the current status and the recent advances in PDAC treatment together with the biological and immunological hallmarks of this cancer entity. On this basis, we discuss new concepts combining distinct treatment modalities in order to improve therapeutic efficacy and clinical outcome – with a specific focus on protocols involving radio(chemo)therapeutic approaches.

 

Pancreatic ductal adenocarcinomas (PDACs), the most prevalent pancreatic cancer, is highly aggressive with a 5-year overall survival rate of less than 8%.  Late detection, it´s metastatic spread and the resistance to chemotherapy are among some of the factors responsible for poor patient outcome. The PDAC incidence rate is increasing and is particularly related to the general aging of our society. Furthermore, life stye habits that include abuse of alcohol and tobacco as well as obesity and type 2 diabetes increases the risk for various types of cancer including PDAC.

 

Do you need support in meeting your research goals? We can help! We offer custom analytical service measurements for ELISA Kits, Luminex Assays, or microRNA Analysis .

GET IN TOUCH at  Biomedica for your custom analytical testing service.

Custom Analytical Service Measurements for ELISA, Luminex, microRNA

Why choose Biomedica Services?

• Expertise – experienced laboratory staff
• Quality – highest quality equipment
• Flexibility – customized according to your project needs and budget
• Speed – rapid turn-around time to meet your deadlines
• Results – verified and comprehensive results presented in an analytical report

Click here for detailed information on our Service Measurements 

RELATED PUBLICATIONS

Comprehensive Characterization of Platelet-Enriched MicroRNAs as Biomarkers of Platelet Activation.

Krammer TL, Zeibig S, Schrottmaier WC, Pirabe A, Goebel S, Diendorfer AB, Holthoff HP, Assinger A, Hackl M. Cells. 2022 Apr 7;11(8):1254. doi: 10.3390/cells11081254. PMID: 35455934; PMCID: PMC9030873.

Abstract

Dysregulation of platelet function is causally connected to thrombus formation and cardiovascular diseases. Therefore, assessing platelet reactivity is crucial. However, current platelet function tests come with pitfalls, limiting clinical use. Plasma miRNA signatures have been suggested as novel biomarkers for predicting/diagnosing cardiovascular diseases and monitoring antiplatelet therapy. Here, we provide results from a comprehensive study on the feasibility of using circulatory platelet miRNAs as surrogate markers of platelet activation. We performed small RNA-Seq on different blood cell types to confirm known and identify novel platelet-enriched miRNAs and validated a panel of 16 miRNAs using RT-qPCR. To identify the main carrier of these blood-based platelet miRNAs, we enriched and analyzed distinct microvesicle populations. Platelets were stimulated with GPVI and P2Y12 agonists in vitro to monitor the release of the selected miRNAs following activation. Finally, the miRNA panel was also measured in plasma from mice undergoing the Folts intervention (recurrent thrombus formation in the carotid artery). Applying an unbiased bioinformatics-supported workflow to our NGS data, we were able to confirm a panel of previously established miRNA biomarker candidates and identify three new candidates (i.e., miR-199a-3p, miR-151a-5p, and miR-148b-3p). Basal levels of platelet-derived miRNAs in plasma were mainly complexed with proteins, not extracellular vesicles. We show that changes in miRNA levels due to platelet activation are detectable using RT-qPCR. In addition, we highlight limitations of studying the in vitro release of miRNAs from platelets. In vivo thrombosis resulted in significant elevations of platelet-derived miRNA levels in mice. In conclusion, we provide in-depth evidence that activated platelets release miRNAs, resulting in measurable changes in circulatory miRNA levels, rendering them promising biomarker candidates.

Multiplex Soluble Biomarker Analysis from Pleural Effusion.

Biomolecules. Javadi J, Dobra K, Hjerpe A. 2020 Jul 28;10(8):1113. doi: 10.3390/biom10081113. PMID: 32731396; PMCID: PMC7464384.

Abstract

Malignant pleural mesothelioma (MPM) is a highly aggressive and therapy resistant pleural malignancy that is caused by asbestos exposure. MPM is associated with poor prognosis and a short patient survival. The survival time is strongly influenced by the subtype of the tumor. Dyspnea and accumulation of pleural effusion in the pleural cavity are common symptoms of MPM. The diagnostic distinction from other malignancies and reactive conditions is done using histopathology or cytopathology, always supported by immunohistochemistry, and sometimes also by analyses of soluble biomarkers in effusion supernatant. We evaluated the soluble angiogenesis related molecules as possible prognostic and diagnostic biomarkers for MPM by Luminex multiplex assay. Pleural effusion from 42 patients with malignant pleural mesothelioma (MPM), 36 patients with adenocarcinoma (AD) and 40 benign (BE) effusions were analyzed for 10 different analytes that, in previous studies, were associated with angiogenesis, consisting of Angiopoietin-1, HGF, MMP-7, Osteopontin, TIMP-1, Galectin, Mesothelin, NRG1-b1, Syndecan-1 (SDC-1) and VEGF by a Human Premixed Multi-Analyte Luminex kit. We found that shed SDC-1 and MMP-7 levels were significantly lower, whereas Mesothelin and Galectin-1 levels were significantly higher in malignant mesothelioma effusions, compared to adenocarcinoma. Galectin-1, HGF, Mesothelin, MMP-7, Osteopontin, shed SDC-1, NRG1-β1, VEGF and TIMP-1 were significantly higher in malignant pleural mesothelioma effusions compared to benign samples. Moreover, there is a negative correlation between Mesothelin and shed SDC-1 and positive correlation between VEGF, Angiopoietin-1 and shed SDC-1 level in the pleural effusion from malignant cases. Shed SDC-1 and VEGF have a prognostic value in malignant mesothelioma patients. Collectively, our data suggest that MMP-7, shed SDC-1, Mesothelin and Galectin-1 can be diagnostic and VEGF and SDC-1 prognostic markers in MPM patients. Additionally, Galectin-1, HGF, Mesothelin, MMP-7, Osteopontin, shed SDC-1 and TIMP-1 can be diagnostic for malignant cases.

Biomarkers & Bone Health – ELISA Kits for Clinical Research

Bone remodeling is a continuous process that removes bone and replaces it with newly synthesized bone. This bone turnover process preserves the mechanical function of the human skeleton.

Bone turnover biomarkers, e.g. markers of bone formation and bone resorption, have been used during the last decade to monitor bone diseases and to monitor their treatment.

Many of these markers are secreted by osteoblasts and osteoclasts and include regulators of bone turnover e.g. receptor activator of NF-kB ligand (RANKL) and osteoprotegerin (OPG).  

Though RANKL and OPG  play an integral role in bone turnover, they do not reflect the activity of osteocytes, the most abundant cell type in the bone.

Osteocytes are cells that regulate bone remodeling. They secrete proteins – bone regulation markers – that include Sclerostin (SOST), Dickkopf-1 (DKK-1), and Fibroblast growth factor  (FGF23). These markers reflect the osteocyte activity.

The above listed biomarkers circulate and can be measured in serum and plasma allowing the investigation of complex interactions between the bone and their relationship with other organs.

BIOMEDICA ELISA KITS – Biomarkers & Bone Health

check out our Bone Biomarker Brochure

ELISA Assay Kit Highlights

+ EASY – ready to use calibrators & controls included (color-coded reagents)
+FULL VALIDATION PACKAGE – assays are optimized for clinical samples
+ HIGH QUALITY GUARANTEED – results you can rely on
+ WIDELY CITED in 1500 + publications

Biomedica – Complete ready-to-use ELISA kits for superior performance and reproducibility

RELATED PRODUCTS

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

FGF23 ELISA , IL-6 ELISA  ,  VEGF ELISA ,  Angiopoietin-2 ELISA

MICRORNA  Analysis 

osteomiR^®– bone miRNA biomarkers highlights:

• Novel minimally invasive biomarkers to detect high imminent fracture-risk in osteoporosis
• 19 individual bone-related biomarkers with distinct information content

RELATED PUBLICATIONS

Novel biological markers of bone: from bone metabolism to bone physiology. Rheumatology (Oxford). Chapurlat RD, Confavreux CB. 2016; 55(10):1714-25.

Abstract

Biochemical markers of bone turnover have been used for decades in the management of bone diseases, to assess the prognosis of these conditions and to monitor treatments. The new markers, however, also reflect specific physiological mechanisms in the bone or other organs. Periostin may be more specific to the periosteum; cathepsin K is an osteoclastic enzyme that may be involved in the cardiovascular system and joints; Dickkopf-1 is involved in bone formation and vascular calcification; sclerostin is a major regulator of bone formation in response to mechanical loading and may also play a role in chronic kidney disease bone and mineral disorder; sphingosine-1-phosphate is a lipid mediator interacting with bone resorption. Some of the bone markers are in fact hormones produced by the bone that affect various physiological and pathological functions in other organs. Thus, osteocalcin is produced by osteoblasts and participates in the regulation of insulin sensitivity and fertility in men. Fibroblast growth factor 23 is produced by osteocytes to regulate phosphorus and 1,25(OH)2D3, but it also plays a major role in the adverse consequences of declining renal function, in particular with respect to the myocardium. Micro RNAs are single-stranded RNAs that regulate several pathways, including the development timing, organogenesis, cell apoptosis, proliferation and differentiation. Their serum concentration may reflect the links between bone physiology and certain conditions in other organs, for example, the cardiovascular system.

Bones have many important biological functions. Bone biomarkers have gained attention in clinical research for the assessment of bone-related diseases. Some of the biomarker proteins have been found to represent useful targets for therapeutic antibodies.

Biomarkers in Bone Biology

Function of the human skeleton

The human skeletal system gives the body it´s structure and helps to protect and support the internal organs. It forms a part of the muscular-skeletal systems that helps the body to move. Throughout our lifetime, the human skeleton undergoes constant remodeling. This dynamic process, degrading bone and replacing it with new tissue maintains bone mass. The continuous cycle of bone resorption and bone growth is also known as bone metabolism.

Bone remodeling

Bone remodeling is a tightly regulated process performed by hormones, cell-signaling molecules, and bone cells. These specific bone cells are osteoclasts, osteoblasts, and osteocytes. The cells are in constant communication with each other through secreted factors, such as osteoprotegerin, RANKL, and sclerostin. These regulatory systems keep the bone remodeling balanced. Imbalances in bone metabolism can lead to bone diseases.

Role of RANKL, RANK, and OPG in bone biology

 Bones are broken down by osteoclasts and rebuilt by osteoblasts.

RANKL receptor activator is a mediator of bone resorption and OPG acts as a decoy receptor.

 

Osteoprotegerin (OPG) is produced by osteoblasts, cells that synthesize bone. OPG is a decoy receptor and binds to RANKL, antagonizing its binding to RANK.

RANKL (receptor activator of nuclear factor kappa-B ligand) is secreted by osteoblasts and binds to the RANK receptor on osteoclast precursor and mature osteoclast cells. RANKL stimulates bone resorption.

 

Role of Sclerostin, FGF23, DKK-1, and Periostin in bone biology

 

 

Biomarkers in Bone Biology

 

Bone cells have been reported to have endocrine functions that affect multiple organs. The most abundant cell type in the bone are osteocytes residing within the bone matrix and comprising 90% to 95% of the bone cells. Osteocytes play a significant role in the regulation of osteogenesis, releasing osteocyte-related biomarkers such as sclerostin (SOST), fibroblast growth factor 23 (FGF23), and Dickkopf-1 (DKK-1).

Sclerostin (SOST) is mainly produced by osteocytes and is considered as the major regulator of bone formation. More recently, Sclerostin has been shown to stimulate the osteocyte synthesis of fibroblast growth factor-23, potentially contributing to the regulation of phosphate homeostasis.

Fibroblast growth factor 23 (FGF23) is a hormone that is mainly secreted by osteocytes and osteoblasts. It regulates phosphate and vitamin D levels and functions as a central endocrine hormone regulating phosphate balance.

Dickkopf-1 (DKK-1) is an extracellular protein. DKK-1 plays a role in the regulation of bone metabolism, as it inhibits the differentiation of osteoblasts.

Periostin (POSTN) is an extracellular matrix protein that is preferentially expressed in the periosteum, a membrane covering the outer surface of bones which is responsible for growth.  Periostin has functions in osteology, tissue repair, oncology, cardiovascular and respiratory diseases, and in a variety of  inflammatory settings (e.g. asthma).

BIOMEDICA OFFERS HIGH QUALITY ELISA KITS FOR BONE BIOMARKERS 

 

widely cited in over 1100 publications

kit validations follow international quality guidelines

 

Sclerostin ELISA (cat.no. BI-20492)

OPG ELISA (cat.no. BI-20403)

RANKL ELISA (cat.no. BI-20462)

FGF23 intact ELISA (cat.no. BI-20700)

FGF23 C-terminal ELISA (cat.no. BI-20702)

DKK-1 ELISA (cat.no. BI-20413)

PERIOSTIN ELISA (cat.no. BI-20433)

 

RELATED LITERATURE

A Review of the Potential Application of Osteocyte-Related Biomarkers, Fibroblast Growth Factor-23, Sclerostin, and Dickkopf-1 in Predicting Osteoporosis and Fractures. Ramli FF, Chin KY. Diagnostics (Basel). 2020 Mar 6;10(3):145. doi: 10.3390/diagnostics10030145. PMID: 32155909; PMCID: PMC7151094.

Sclerostin Directly Stimulates Osteocyte Synthesis of Fibroblast Growth Factor-23. Wijenayaka AR, Yang D, Ormsby RT, Bonewald LF, Atkins GJ. Calcif Tissue Int. 2021 Jul;109(1):66-76. doi: 10.1007/s00223-021-00823-6. Epub 2021 Feb 22. PMID: 33616712.

Biology of the RANKL-RANK-OPG System in Immunity, Bone, and Beyond. Front Immunol. Walsh MC, Choi Y.2014 Oct 20;5:511. doi: 10.3389/fimmu.2014.00511. PMID: 25368616; PMCID: PMC4202272.

The Osteocyte: New Insights. Robling AG, Bonewald LF. Annu Rev Physiol. 2020 Feb 10;82:485-506. doi: 10.1146/annurev-physiol-021119-034332. PMID: 32040934; PMCID: PMC8274561.

 

 

Sema4D induces cartilage destruction

Semaphorin 4D destroys cartilage: Semaphorin 4D (Sema 4D) has been identified as an inflammatory cytokine that promotes cartilage destruction. This finding was recently published by Murakami T and colleagues. In a mouse model of inflammatory arthritis, the researchers demonstrated that Sema4D was increased in synovial fluid and loss of Sema4D protected against cartilage degeneration. Click here to learn more.

 

 

What is cartilage?

Cartilage is a type of strong and flexible tissue that is found throughout the body. It is a connective tissue that exists in our joints, bones, spine, ears and nose. It protects our joints and our bones, absorbing impact, reducing friction and helping our joints to move smoothly.

 If cartilage is degraded, movements will cause friction and pain, leading to swelling and stiffness. Cartilage degradation can be caused through direct injury, osteoarthritis, and aging.  

Semaphorin 4D destroys cartilage

Semaphorin 4D induces articular cartilage destruction and inflammation in joints by transcriptionally reprogramming chondrocytes.

Murakami T, Takahata Y, Hata K, Ebina K, Hirose K, Ruengsinpinya L, Nakaminami Y, Etani Y, Kobayashi S, Maruyama T, Nakano H, Kaneko T, Toyosawa S, Asahara H, Nishimura R. Sci Signal. 2022 Nov;15(758):eabl5304. doi: 10.1126/scisignal.abl5304. Epub 2022 Nov 1. PMID: 36318619.

Abstract

Proinflammatory cytokines play critical roles in the pathogenesis of joint diseases. Using a mass spectrometry-based cloning approach, we identified Semaphorin 4D (Sema4D) as an inflammatory cytokine that directly promoted cartilage destruction. Sema4d-deficient mice showed less cartilage destruction than wild-type mice in a model of rheumatoid arthritis. Sema4D induced a proinflammatory response in mouse articular chondrocytes characterized by the induction of proteolytic enzymes that degrade cartilage, such as matrix metalloproteinases (MMPs) and aggrecanases. The activation of Mmp13 and Mmp3 expression in articular chondrocytes by Sema4D did not depend on RhoA, a GTPase that mediates Sema4D-induced cytoskeletal rearrangements. Instead, it required NF-κB signaling and Ras-MEK-Erk1/2 signaling downstream of the receptors Plexin-B2 and c-Met and depended on the transcription factors IκBζ and C/EBPδ. Genetic and pharmacological blockade of these Sema4D signaling pathways inhibited MMP induction in chondrocytes and cartilage destruction in femoral head organ culture. Our results reveal a mechanism by which Sema4D signaling promotes cartilage destruction.

SEMA4D ELISA kit highlights

First fully validated Sema4D assay (cat.no. BI-20405)

10 µl sample/well

Reference values provided

Developed & manufactured by Biomedica

RELATED PUBLICATIONS

Inhibition of Semaphorin 4D/Plexin-B1 signaling inhibits the subchondral bone loss in early-stage osteoarthritis of the temporomandibular joint. Zhang Z, Lu L, Ye T, Yu S, Zhang J, Zhang M, He F, Liu Q, Yang H, Feng J.Arch Oral Biol. 2022 Mar;135:105365. doi: 10.1016/j.archoralbio.2022.105365. Epub 2022 Feb 2. PMID: 35151027.

Abstract

Compliance of NT-proBNP test in accredited cardiac marker survey

NT-proBNP ELISA test successfully passed accredited cardiac marker survey: Biomedica participated in an accredited international ring trial program for the cardiac biomarker NT-proBNP.  The results demonstrate that the CE-marked NT-proBNP ELISA assay successfully passed the QC circle in the cardiac marker survey.

NT-proBNP ELISA test successfully passed cardiac marker survey

The results comply with strict quality standards confirmed by RfB, an international provider for proficiency testing.

NT-proBNP ELISA assay kit (cat. no. SK-1204)

√  CE-marked – for IVD use in the EU
√  Flexible – can be run in every lab
√  Two controls included
√  Simple 2 step protocol
√  Reliable – full validation
√  Widely cited in more than 100 publications

 

Developed & manufactured by Biomedica  right in the heart of Europe

 

 

•  RfB NT-proBNP Biomedica Certificate Proficiency Testing

 

 

What is proficiency testing?

Proficiency Testing provides a report that shows how laboratories can compare their own data to data to data from other laboratories around the world.

The testing, carried out in a ring trial program,  is performed by an accredited laboratory specialist in proficiency testing. Proficiency testing monitors the performance of individual laboratories for specific tests e.g. cardiac biomarker assays.

LITERATURE

Monitoring of NT-proBNP – predicting risk of cardiovascular events from anti-inflammatory drugs (Non-Steroid Anti-Inflammatory Drugs)

N-terminal pro-B-type natriuretic peptide concentrations predict the risk of cardiovascular adverse events from antiinflammatory drugs: a pilot trial. Brune K, Katus HA, Moecks J, Spanuth E, Jaffe AS, Giannitsis E. Clin Chem. 2008 Jul;54(7):1149-57. doi: 10.1373/clinchem.2007.097428. Epub 2008 May 1. PMID: 18451314.

Safety of Oral Non-Selective Non-Steroidal Anti-Inflammatory Drugs in Osteoarthritis: What Does the Literature Say? Cooper C, Chapurlat R, Al-Daghri N, Herrero-Beaumont G, Bruyère O, Rannou F, Roth R, Uebelhart D, Reginster JY.Drugs Aging. 2019 Apr;36(Suppl 1):15-24. doi: 10.1007/s40266-019-00660-1. PMID: 31073921; PMCID: PMC6509083.

Cardio-renal safety of non-steroidal anti-inflammatory drugs. Radi ZA, Khan KN. J Toxicol Sci. 2019;44(6):373-391. doi: 10.2131/jts.44.373. PMID: 31168026.

Abstract

Cardiovascular safety of non-steroidal anti-inflammatory drugs: network meta-analysis. Trelle S, Reichenbach S, Wandel S, Hildebrand P, Tschannen B, Villiger PM, Egger M, Jüni P.BMJ. 2011 Jan 11;342:c7086. doi: 10.1136/bmj.c7086. PMID: 21224324; PMCID: PMC3019238.

 

Pathogenic contribution of LRG1 to vascular retinopathies  

Globally, 2.2 billion people are estimated to have a vision impairment. Eye diseases are on the rise due to an ageing population and an increase of diabetes. The classical therapy involves the blockage of vascular endothelial growth factor (VEGF). However, a high rate of patient non-response and loss of efficacy over time are major challenges.

Evaluating new treatments with better efficiency are moving forward. One novel therapeutic target is leucine-rich α-2 glycoprotein 1 (LRG1). It is an  emerging key player in vascular dysfunction, inflammation, and fibrosis.

LRG1 as a novel therapeutic target in eye disease

The uncontrolled formation of new blood vessels in the eye (ocular angiogenesis) is one of the major causes of eye diseases and blindness.

The blood protein LRG1 plays a central role in the progression of eye diseases and is thus a promising novel therapeutic target.

In searching for mediators of vascular remodeling in the diseased and damaged retina, researchers discovered that leucine-rich α-2 glycoprotein 1 (LRG1) is a novel regulator of TGFß signaling. The study published in Nature in 2013 by Wang X et al., revealed that LRG1 is a novel regulator of angiogenesis that mediates its effect through modulating TGFβ signaling. The researchers showed that LRG1 expression in the retina is predominantly vascular and is increased during neovascular growth. Their study in mice supported the hypothesis that LRG1 is necessary for robust vascular growth and its inhibition has potential as a therapeutic target. Based on their studies, the authors suggested that LRG1 is not only a promising target for controlling pathogenic angiogenesis in eye diseases but could potentially also be used in other diseases such as cancer and atherosclerosis.

WHAT IS LRG1 (leucine-rich α-2 glycoprotein 1)

The mature 38,2 kDa glyocoprotein LRG contains 347 amino acids (Uniprot entry Leucine-rich alpha-2-glycoprotein). It belongs to the family of LRG proteins characterized through leucine-rich repeats in its amino acid sequence. LRG1 (also named LRG) is involved in protein-protein interactions, signal transduction, and development.  Studies have shown that LRG plays a role in physiological processes of the nervous system including synapse formation and growth.  It is also implied in cell proliferation, in immune responses, in cell migration, in neovascularization, and in apoptosis.

Alternative titles: LRG – OMIM Entry leucine-rich α-2 glycoprotein (LRG)

HOW CAN YOU MEASURE LRG1?

LRG1 can reliably be measured in human serum and plasma with the Biomedica LRG ELISA (cat. no. BI-LRG). 

• Sample volume: 5µl

• Assay range: 2-64 ng/ml   

•  Easy protocol – day test

 

The validation of the LRG1 ELISA kit has been performed following international quality guidelines. Download the validation data here.

RELATED PUBLICATIONS

LRG1 as a novel therapeutic target in eye disease.

De Rossi G, Da Vitoria Lobo ME, Greenwood J, Moss SE. Eye (Lond). 2022 Feb;36(2):328-340. doi: 10.1038/s41433-021-01807-4. Epub 2022 Jan 5. Erratum in: Eye (Lond). 2022 Feb 28;: PMID: 34987199; PMCID: PMC8807626.

Abstract

Retinal and choroidal diseases are major causes of blindness and visual impairment in the developed world and on the rise due to an ageing population and diabetes epidemic. Standard of care is centred around blockade of vascular endothelial growth factor (VEGF), but despite having halved the number of patients losing sight, a high rate of patient non-response and loss of efficacy over time are key challenges. Dysregulation of vascular homoeostasis, coupled with fibrosis and inflammation, are major culprits driving sight-threatening eye diseases. Improving our knowledge of these pathological processes should inform the development of new drugs to address the current clinical challenges for patients. Leucine-rich α-2 glycoprotein 1 (LRG1) is an emerging key player in vascular dysfunction, inflammation and fibrosis. Under physiological conditions, LRG1 is constitutively expressed by the liver and granulocytes, but little is known about its normal biological function. In pathological scenarios, such as diabetic retinopathy (DR) and neovascular age-related macular degeneration (nvAMD), its expression is ectopically upregulated and it acquires a much better understood pathogenic role. Context-dependent modulation of the transforming growth-factor β (TGFβ) pathway is one of the main activities of LRG1, but additional roles have recently been emerging. This review aims to highlight the clinical and pre-clinical evidence for the pathogenic contribution of LRG1 to vascular retinopathies, as well as extrapolate from other diseases, functions which may be relevant to eye disease. Finally, we will provide a current update on the development of anti-LRG1 therapies for the treatment of nvAMD..

LRG1 promotes angiogenesis by modulating endothelial TGF-β signalling. 

Wang X, Abraham S, McKenzie JAG, Jeffs N, Swire M, Tripathi VB, Luhmann UFO, Lange CAK, Zhai Z, Arthur HM, Bainbridge J, Moss SE, Greenwood J.Nature. 2013 Jul 18;499(7458):306-11. doi: 10.1038/nature12345. PMID: 23868260; PMCID: PMC3836402.

High big ET-1 levels are associated with risk of all–cause death in diabetes patients with CAD

Big Endothelin-1 associated with all-cause death in diabetes patients with CAD

A clinical study in 8550 patients investigated the association between the cardio-vascular marker big endothelin-1 (big ET-1) and long-term all-cause death in patients with coronary artery disease (CAD) and impaired glucose metabolism.

Patients were categorized into both glucose status (diabetes mellitus, pre-diabetes mellitus, normalglycemia) and big ET-1 level groups. Primary endpoint was all-cause death. The data indicate that baseline big ET-1 levels were independently associated with the long-term mortality in diabetes patients with CAD.

Read more Big Endothelin-1 associated with all-cause death in diabetes patients with CAD.

What is Big Endothelin-1 (Big ET-1) ?

Big ET-1 is a 38 amino acid peptide and is the precursor of Endothelin-1 (ET-1). ET-1 is a potent vasoconstrictor secreted by endothelial cells. It acts as the counterpart of the vasodilator nitric oxide (NO).

• ET-1 is cleaved from Big ET-1 by the ET converting enzyme -1 (ECE-1).

• Both the precursor Big ET-1 and the shorter 21 aminoacid vasoactive form Endothelin-1 circulate in blood.

• ET-1, the biologically active form, is rapidly cleared and has a short half-life.

• ET-1 and big ET-1 are found in equimolar concentrations in the plasma.

• Thus, the longer half-life of big ET-1, its slower clearance and the findings that increased plasma levels of ET-1 in patients with heart failure are mainly due to an increase in big ET-1 concentrations, offers an analytical window for the measurement of Big ET-1 .

How can you measure Big ET-1?

Big ET-1 can easily be measured using  in serum and plasma with only 50µl sample volume.

Biomedica’s Big ET-1 ELISA cat. no. BI-20082H

√ Direct measurement – highly sensitive
√ Full validation package
√ Widely cited +75 publications

check out the customer review on Biocompare 

Related publications – Citations Big ET-1 ELISA – Biomedica

The Biomedica Big Endothelin-1 (Big ET-1) ELISA kit was utilized in the following studies:

Big Endothelin-1 and long-term all-cause death in patients with coronary artery disease and prediabetes or diabetes after percutaneous coronary intervention

Xu N, Zhu P, Yao Y, Jiang L, Jia S, Yuan D, Xu J, Wang H, Song Y, Gao L, Gao Z, Song L, Zhao X, Chen J, Yang Y, Xu B, Gao R, Yuan J. Nutr Metab Cardiovasc Dis. 2022 Sep;32(9):2147-2156. doi: 10.1016/j.numecd.2022.06.002. Epub 2022 Jun 11. PMID: 35843800.

Abstract

Background and aims: The present study aimed to examine the association between big endothelin-1 (big ET-1) and long-term all-cause death in patients with coronary artery disease (CAD) and different glucose metabolism status.

Methods and results: We consecutively enrolled 8550 patients from January 2013 to December 2013. Patients were categorized according to both status of glucose metabolism status [Diabetes Mellitus (DM), Pre-Diabetes (Pre-DM), Normoglycemia (NG)] and big ET-1 levels. Primary endpoint was all-cause death. During a median of 5.1-year follow-up periods, 301 all-cause deaths occurred. Elevated big ET-1 was significantly associated with long-term all-cause death (adjusted HR: 2.230, 95%CI 1.629-3.051; p < 0.001). Similarly, patients with DM, but not Pre-DM, had increased risk of all-cause death compared with NG group (p < 0.05). When patients were categorized by both status of glucose metabolism and big ET-1 levels, high big ET-1 were associated with significantly higher risk of all-cause death in Pre-DM (adjusted HR: 2.442, 95% CI 1.039-5.740; p = 0.041) and DM (adjusted HR: 3.162, 95% CI 1.376-7.269; p = 0.007). The Kaplan-Meier curve indicated that DM patients with the highest big ET-1 levels were associated with the greatest risk of all-cause death (p < 0.05).

Conclusions: The present data indicate that baseline big ET-1 levels were independently associated with the long-term all-cause death in DM and Pre-DM patients with CAD undergoing PCI, suggesting that big ET-1 may be a valuable marker in patients with impaired glucose metabolism.

Predictive Value of Plasma Big Endothelin-1 in Adverse Events of Patients With Coronary Artery Restenosis and Diabetes Mellitus: Beyond Traditional and Angiographic Risk Factors. Ma Y, Tian T, Wang T, Wang J, Guan H, Yuan J, Song L, Yang W, Qiao S. Front Cardiovasc Med. 2022 May 26;9:854107. doi: 10.3389/fcvm.2022.854107. PMID: 35694656; PMCID: PMC9177997.

Big Endothelin-1 and long-term all-cause death in patients with coronary artery disease and prediabetes or diabetes after percutaneous coronary intervention . Xu N, Zhu P, Yao Y, Jiang L, Jia S, Yuan D, Xu J, Wang H, Song Y, Gao L, Gao Z, Song L, Zhao X, Chen J, Yang Y, Xu B, Gao R, Yuan J. Nutr Metab Cardiovasc Dis. 2022 Sep;32(9):2147-2156. doi: 10.1016/j.numecd.2022.06.002. Epub 2022 Jun 11. PMID: 35843800.

Plasma concentration of big endothelin-1 and its relation with plasma NT-proBNP and ventricular function in heart failure patients. Rivera M, Cortés R, Portolés M, Valero R, Sancho-Tello MJ, Martínez-Dolz L, Sevilla B, Taléns-Visconti R, Jordán A, Miró V, Pérez-Boscá JL, Marín F, Climent V, García de Burgos F, Payá R, Sogorb F, Bertomeu V, Salvador A. Rev Esp Cardiol. 2005 Mar;58(3):278-84. Spanish. PMID: 15766450.

Plasma big endothelin-1 concentrations in congestive heart failure patients with or without systemic hypertension. Pacher R, Bergler-Klein J, Globits S, Teufelsbauer H, Schuller M, Krauter A, Ogris E, Rödler S, Wutte M, Hartter E. Am J Cardiol. 1993 Jun 1;71(15):1293-9. doi: 10.1016/0002-9149(93)90543-l. PMID: 8498369.

Plasma big endothelin-1 levels at admission and future cardiovascular outcomes: A cohort study in patients with stable coronary artery disease. Zhou BY, Guo YL, Wu NQ, Zhu CG, Gao Y, Qing P, Li XL, Wang Y, Dong Q, Liu G, Xu RX, Cui CJ, Sun J, Li JJ. Int J Cardiol. 2017 Mar 1;230:76-79. doi: 10.1016/j.ijcard.2016.12.082. Epub 2016 Dec 21. PMID: 28038820.

Background: Big endothelin-1 (ET-1) has been proposed as a novel prognostic indicator of acute coronary syndrome, while its predicting role of cardiovascular outcomes in patients with stable coronary artery disease (CAD) is unclear.

Methods and results: A total of 3154 consecutive patients with stable CAD were enrolled and followed up for 24months. The outcomes included all-cause death, non-fatal myocardial infarction, stroke and unplanned revascularization (percutaneous coronary intervention and coronary artery bypass grafting). Baseline big ET-1 was measured using sandwich enzyme immunoassay method. Cox proportional hazard regression analysis and Kaplan-Meier analysis were used to evaluate the prognostic value of big ET-1 on cardiovascular outcomes. One hundred and eighty-nine (5.99%) events occurred during follow-up. Patients were divided into two groups: events group (n=189) and non-events group (n=2965). The results indicated that the events group had higher levels of big ET-1 compared to non-events group. Multivariable Cox proportional hazard regression analysis showed that big ET-1 was positively and statistically correlated with clinical outcomes (Hazard Ratio: 1.656, 95% confidence interval: 1.099-2.496, p=0.016). Additionally, the Kaplan-Meier analysis revealed that patients with higher big ET-1 presented lower event-free survival (p=0.016).

Conclusions: The present study firstly suggests that big ET-1 is an independent risk marker of cardiovascular outcomes in patients with stable CAD. And more studies are needed to confirm our findings.

Superiority of big endothelin-1 and endothelin-1 over natriuretic peptides in predicting survival in severe congestive heart failure: a 7-year follow-up study. Van Beneden R, Gurné O, Selvais PL, Ahn SA, Robert AR, Ketelslegers JM, Pouleur HG, Rousseau MF. J Card Fail. 2004 Dec;10(6):490-5. doi: 10.1016/j.cardfail.2004.04.001. PMID: 15599839.

Try Biomedica’s Cytokine ELISA Kits for free!

Annoyed wasting valuable resources by inefficient tests?

Order your free trial Cytokine ELISA kit – offer valid until 12/31/22.  

Contact us at Biomedica: info@bmgrp.com

Discover Biomedica’s Cytokine Kits – Use code: Biomedica FREE CytELISA Promotion – CN040010:

Discover Biomedica’s Cytokine Kits

 

Full validation package- assays are optimized for clinical samples

human Cytokine ELISA Kits  

developed & manufactured by Biomedica.  Austrian Quality.

IL-6 (Interleukin-6)             high sensitivity
VEGF                                           low sample volume – 10µl
ANGIOPOIETIN-2              optimized assay range

 All kits include color coded, ready to use prediluted standards and controls.

 Citations

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. Nephrology Dialysis Transplantation, Volume 34, Issue Supplement_1, June 2019, gfz103.SP339, https://doi.org/10.1093/ndt/gfz103.SP339

Click here to view poster: ERA-EDTA Poster 2019 Human Angiopoietin-2

 

Abstract

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.

RELATED LITERATURE 

1. Molnar et al. (2014): Circulating angiopoietin-2 levels predict mortality in kidney transplant recipients: a 4-year prospective case–cohort study. Transplant International, 27 (6): 541-52. 2. David et al. (2010): Circulating angiopoietin-2 levels increase with progress of chronic kidney disease. Nephrol. Dial. Transplant, 25: 2571-2579,
2. Tsai et al. (2017): The interaction between fluid status and angiopoietin-2 in adverse renal outcomes of chronic kidney disease. PLoS One, 12 (3): e173906.
3. Tsai et al. (2016): Angiopoietin-2, Angiopoietin-1 and subclinical cardiovascular disease in Chronic Kidney Disease. Scientific Reports, 6:39400

 

Circulating DKK-1 levels predict disease outcomes and mirror metabolic adaptations in patients with Covid-19

Individuals with low serum levels of DKK1 (Dickkopf-1) are twice as likely to die from Covid-19 than those with high levels according to new research published by Nikolai Jaschke and colleagues in the Journal of Clinical Endocrinology & Metabolism.

The researchers found that circulating DKK1 levels vary in humans and change as a function of time during SARS-CoV-2 infection. The infection promotes metabolic adaptations that resembles fasting, which are mirrored by circulating DKK1 levels.  DKK-1 levels predict disease outcomes in Covid-19 individuals.

The results of the study suggest a potential clinical use of measuring circulating DKK1 as an indicator of disease severity in COVID-19 patients. 

Read more: Circulating Dickkopf1 parallels metabolic adaptations and predicts disease trajectories in patients with Covid-19.

Circulating DKK1 levels were measured  with the DKK-1 ELISA kit from Biomedica

Biomedica, Vienna, Austria, DKK-1 ELISA, catalog #BI-20413, RRID: AB_2922680 

Highlights – DKK-1 ELISA (cat. no. BI-20413)

• Small sample volume – 20µl serum/well
• Reliable –international validation guidelines
• Easy – direct measurement
• Widely cited in +170 publications

 

Circulating Dickkopf1 parallels metabolic adaptations and predicts disease trajectories in patients with Covid-19.   Full text link

Jaschke NP, Funk AM, Jonas S, Riffel RM, Sinha A, Wang A, Pählig S, Hofmann M, Altmann H, Von Bonin S, Koch T, Spieth P, Tausche K, Akgün K, Rauner M, Kronstein-Wiedemann R, Odendahl M, Tonn T, Göbel A, Hofbauer LC, Rachner TD. J Clin Endocrinol Metab. 2022 Sep 8:dgac514. doi: 10.1210/clinem/dgac514. Epub ahead of print. PMID: 36071553; PMCID: PMC9494396.

Abstract

Context and aims: Coronavirus disease 19 (COVID-19) trajectories show high interindividual variability, ranging from asymptomatic manifestations to fatal outcomes, the latter of which may be fueled by immunometabolic maladaptation of the host. Reliable identification of patients who are at risk of severe disease remains challenging. We hypothesized that serum concentrations of Dickkopf1 (DKK1) indicate disease outcomes in severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)-infected individuals.

Methods: We recruited hospitalized patients with PCR-confirmed SARS-CoV-2 infection and included 80 individuals for whom blood samples from 2 independent time points were available. DKK1 serum concentrations were measured by ELISA in paired samples. Clinical data were extracted from patient charts and correlated with DKK1 levels. Publicly available datasets were screened for changes in cellular DKK1 expression on SARS-CoV-2 infection. Plasma metabolites were profiled by nuclear magnetic resonance spectroscopy in an unbiased fashion and correlated with DKK1 data. Kaplan-Meier and Cox regression analysis were used to investigate the prognostic value of DKK1 levels in the context of COVID-19.

Results: We report that serum levels of DKK1 predict disease outcomes in patients with COVID-19. Circulating DKK1 concentrations are characterized by high interindividual variability and change as a function of time during SARS-CoV-2 infection, which is linked to platelet counts. We further find that the metabolic signature associated with SARS-CoV-2 infection resembles fasting metabolism and is mirrored by circulating DKK1 abundance. Patients with low DKK1 levels are twice as likely to die from COVID-19 than those with high levels, and DKK1 predicts mortality independent of markers of inflammation, renal function, and platelet numbers.

Conclusion: Our study suggests a potential clinical use of circulating DKK1 as a predictor of disease outcomes in patients with COVID-19. These results require validation in additional cohorts.

Decrease of bone biomarker Sclerostin in women with anorexia nervosa during nutrition therapy – indication of reduced bone loss

Anorexia nervosa (AN) is an eating disorder and has one of the highest mortality rates of any mental illness.  It affects roughly 2.9 million people and many experience bone loss and increased fracture risk. In a 3-year prospective study, Swedish researchers looked into the long-term effects of nutrition therapy. They  investigated bone and mineral metabolism and biomarkers young women with AN. Their results showed that body mass index (BMI) and fat mass was increased. The regulatory bone biomarker Sclerostin decreased during nutrition therapy and further over 3 years, indicating reduced bone loss.

Read more: Bone mass and biomarkers in young women with anorexia nervosa: a prospective 3-year follow-up study.

Biomedica Sclerostin ELISA kit 

• The internationally most referenced Sclerostin ELISA with more than 270 citations!
• Rigorously validated according to international quality guidelines
• Low sample volume

 

Bone mass and biomarkers in young women with anorexia nervosa: a prospective 3-year follow-up study.

Svedlund A, Pettersson C, Tubic B, Ellegård L, Elfvin A, Magnusson P, Swolin-Eide D. J Bone Miner Metab. 2022 Aug 12. doi: 10.1007/s00774-022-01359-x. Epub ahead of print. PMID: 35960382.

Abstract

Introduction: Anorexia nervosa (AN) increases the risk of impaired bone health, low areal bone mineral density (aBMD), and subsequent fractures. This prospective study investigated the long-term effects of bone and mineral metabolism on bone and biomarkers in 22 women with AN.

Materials and methods: Body composition and aBMD were measured by dual-energy X-ray absorptiometry (DXA) and peripheral quantitative computed tomography. Total and free 25-hydroxyvitamin D (25OHD), C-terminal collagen cross-links (CTX), osteocalcin, bone-specific alkaline phosphatase (BALP), leptin, sclerostin, and oxidized/non-oxidized parathyroid hormone (PTH) were analyzed before and after 12 weeks of intensive nutrition therapy and again 3 years later. An age-matched comparison group of 17 healthy women was recruited for the 3-year follow-up.

Results: Body mass index (BMI) and fat mass increased from baseline to 3 years in women with AN. Sclerostin decreased during nutrition therapy and further over 3 years, indicating reduced bone loss. CTX was elevated at baseline and after 12 weeks but decreased over 3 years. BALP increased during nutrition therapy and stabilized over 3 years. Free 25OHD was stable during treatment but decreased over 3 years. Non-oxidized PTH was stable during treatment but increased over 3 years. Trabecular volumetric BMD in AN patients decreased during the first 12 weeks and over 3 years despite stable BMI and bone biomarkers implying increased BMD.

Conclusion: Our findings highlight the importance of early detection and organized long-term follow-up of bone health in young women with a history of AN.

Keywords: DXA; Eating disorder; Osteoporosis; Sclerostin; Vitamin D

Related publications

Anorexia nervosa: 30-year outcome.

Dobrescu SR, Dinkler L, Gillberg C, Råstam M, Gillberg C, Wentz E. Br J Psychiatry. 2020 Feb;216(2):97-104. doi: 10.1192/bjp.2019.113. PMID: 31113504; PMCID: PMC7557598.

Abstract

Background: Little is known about the long-term outcome of anorexia nervosa.

Aims: To study the 30-year outcome of adolescent-onset anorexia nervosa. 

Method: All 4291 individuals born in 1970 and attending eighth grade in 1985 in Gothenburg, Sweden were screened for anorexia nervosa. A total of 24 individuals (age cohort for anorexia nervosa) were pooled with 27 individuals with anorexia nervosa (identified through community screening) who were born in 1969 and 1971-1974. The 51 individuals with anorexia nervosa and 51 school- and gender-matched controls were followed prospectively and examined at mean ages of 16, 21, 24, 32 and 44. Psychiatric disorders, health-related quality of life and general outcome were assessed. 

Results: At the 30-year follow-up 96% of participants agreed to participate. There was no mortality. Of the participants, 19% had an eating disorder diagnosis (6% anorexia nervosa, 2% binge-eating disorder, 11% other specified feeding or eating disorder); 38% had other psychiatric diagnoses; and 64% had full eating disorder symptom recovery, i.e. free of all eating disorder criteria for 6 consecutive months. During the elapsed 30 years, participants had an eating disorder for 10 years, on average, and 23% did not receive psychiatric treatment. Good outcome was predicted by later age at onset among individuals with adolescent-onset anorexia nervosa and premorbid perfectionism.

Conclusions: This long-term follow-up study reflects the course of adolescent-onset anorexia nervosa and has shown a favourable outcome regarding mortality and full symptom recovery. However, one in five had a chronic eating disorder.

Anorexia Nervosa.

Mitchell JE, Peterson CB. N Engl J Med. 2020 Apr 2;382(14):1343-1351. doi: 10.1056/NEJMcp1803175. PMID: 32242359.

Osteoporosis Awareness – 

Worldwide around 200 million women have osteoporosis. Osteoporosis is a disease that weakens bone. Bones become fragile and porous.  From the outside, the osteoporotic bone is shaped like normal bone. However, the inside of the bone loses density and becomes weak. The risk of fractures become greater with age, due to the loss of minerals like calcium and phosphate. Osteoporosis most often affects bones in the hip, the spine and the wrist. Factors that influence bone health are nutrition, exercise and hormonal factors. Osteoporosis cannot be cured but delayed through early intervention and biomarker research has identified proteins that may help to identify patients at risk.

 

BIOMEDICA offers ELISA kits to measure Bone Biomarkers in human serum or plasma samples:

 

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

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

 How can you prevent Osteoporosis?

Osteoporosis prevention includes a balanced diet, containing sufficient amounts of calcium and vitamin D and regular exercise.

Exercises to protect or reduce your chance of fracture include regular weight-bearing exercise (eg. weight lifting or resistance training) or any kind of activity that carries your own body weight (e.g. walking, running, climbing stairs, dancing).

A healthy and balanced diet with the recommended daily amounts of calcium are important for bone health. Read more about “Good for your bone foods .

Related literature

 The Effectiveness of Physical Exercise on Bone Density in Osteoporotic Patients.

Benedetti MG, Furlini G, Zati A, Letizia Mauro G. Biomed Res Int. 2018. 23;2018:4840531. PMID: 30671455. Link to full text

Abstract

Physical exercise is considered an effective means to stimulate bone osteogenesis in osteoporotic patients. The authors reviewed the current literature to define the most appropriate features of exercise for increasing bone density in osteoporotic patients. Two types emerged: (1) weight-bearing aerobic exercises, i.e., walking, stair climbing, jogging, and Tai Chi. Walking alone did not appear to improve bone mass; however it is able to limit its progressive loss. In fact, in order for the weight-bearing exercises to be effective, they must reach the mechanical intensity useful to determine an important ground reaction force. (2) Strength and resistance exercises: these are carried out with loading (lifting weights) or without (swimming, cycling). For this type of exercise to be effective a joint reaction force superior to common daily activity with sensitive muscle strengthening must be determined. These exercises appear extremely site-specific, able to increase muscle mass and BMD only in the stimulated body regions. Other suggested protocols are multicomponent exercises and whole body vibration. Multicomponent exercises consist of a combination of different methods (aerobics, strengthening, progressive resistance, balancing, and dancing) aimed at increasing or preserving bone mass. These exercises seem particularly indicated in deteriorating elderly patients, often not able to perform exercises of pure reinforcement. However, for these protocols to be effective they must always contain a proportion of strengthening and resistance exercises. Given the variability of the protocols and outcome measures, the results of these methods are difficult to quantify. Training with whole body vibration (WBV): these exercises are performed with dedicated devices, and while it seems they have effect on enhancing muscle strength, controversial findings on improvement of BMD were reported. WBV seems to provide good results, especially in improving balance and reducing the risk of falling; in this, WBV appears more efficient than simply walking. Nevertheless, contraindications typical of senility should be taken into account.

 

Exercise for the prevention of osteoporosis in postmenopausal women: an evidence-based guide to the optimal prescription.

Daly RM, Dalla Via J, Duckham RL, Fraser SF, Helge EW.Braz J Phys Ther. 2019.23(2):170-180. PMID: 30503353. Full text link

Abstract

Background: Osteoporosis and related fragility fractures are a global public health problem in which pharmaceutical agents targeting bone mineral density (BMD) are the first line of treatment. However, pharmaceuticals have no effect on improving other key fracture risk factors, including low muscle strength, power and functional capacity, all of which are associated with an increased risk for falls and fracture, independent of BMD. Targeted exercise training is the only strategy that can simultaneously improve multiple skeletal and fall-related risk factors, but it must be appropriately prescribed and tailored to the desired outcome(s) and the specified target group.

Objectives: In this review, we provide an overview of the general principles of training and specific loading characteristics underlying current exercise guidelines for the prevention of osteoporosis, and an update on the latest scientific evidence with regard to the type and dose of exercise shown to positively influence bone mass, structure and strength and reduce fracture risk in postmenopausal women.

The bone metabolism mediators RANK (receptor activator of nuclear factor κB), RANKL (receptor activator of nuclear factor κB ligand) and OPG (osteoprotegerin) have been linked to breast cancer tumorgenesis. The dysregulation of RANK signaling is involved in the pathogenesis of BRCA1 associated breast cancer.

Experimental studies have shown that OPG (the decoy receptor for RANKL) protein levels are significantly lower in BRCA1 mutation carriers, inhibiting RANK signaling. Thus, OPG may serve as a potential biomarker of breast cancer risk.  

Learn more: Delineating the role of osteoprotegerin as a marker of breast cancer risk among women with a BRCA1 mutation. Park SS et al., 2022.

Osteoprotegerin a potential biomarker of breast cancer risk

OPG can reliably be measured by ELISA analysis with only 20µl of serum or plasma.

The Biomedica OPG ELISA is the most referenced human OPG ELISA.

• Widely cited in more than 240 publications

 

Delineating the role of osteoprotegerin as a marker of breast cancer risk among women with a BRCA1 mutation.

Park SS, Uzelac A, Kotsopoulos J Hered Cancer Clin Pract. 2022 Apr 13;20(1):14. doi: 10.1186/s13053-022-00223-3. PMID: 35418083; PMCID: PMC9008947. Read full publication

 Abstract

Women with a pathogenic germline mutation in the BRCA1 gene face a very high lifetime risk of developing breast cancer, estimated at 72% by age 80. Prophylactic bilateral mastectomy is the only effective way to lower their risk; however, most women with a mutation opt for intensive screening with annual MRI and mammography. Given that the BRCA1 gene was identified over 20 years ago, there is a need to identify a novel non-surgical approach to hereditary breast cancer prevention. Here, we provide a review of the emerging preclinical and epidemiologic evidence implicating the dysregulation of progesterone-mediated receptor activator of nuclear factor κB (RANK) signaling in the pathogenesis of BRCA1-associated breast cancer. Experimental studies have demonstrated that RANK inhibition suppresses Brca1-mammary tumorigenesis, suggesting a potential target for prevention. Data from studies conducted among women with a BRCA1 mutation further support this pathway in BRCA1-associated breast cancer development. Progesterone-containing (but not estrogen-alone) hormone replacement therapy is associated with an increased risk of breast cancer in women with a BRCA1 mutation. Furthermore, BRCA1 mutation carriers have significantly lower levels of circulating osteoprotegerin (OPG), the decoy receptor for RANK-ligand (RANKL) and thus endogenous inhibitor of RANK signaling. OPG levels may be associated with the risk of disease, suggesting a role of this protein as a potential biomarker of breast cancer risk. This may improve upon current risk prediction models, stratifying women at the highest risk of developing the disease, and further identify those who may be targets for anti-RANKL chemoprevention. Collectively, the evidence supports therapeutic inhibition of the RANK pathway for the primary prevention of BRCA1-associated breast cancer, which may generate unique prevention strategies (without prophylactic surgery) and enhance quality of life.

Related publications

Plasma osteoprotegerin and breast cancer risk in BRCA1 and BRCA2 mutation carriers.

Odén L, Akbari M, Zaman T, Singer CF, Sun P, Narod SA, Salmena L, Kotsopoulos J. Oncotarget. 2016.  27;7(52):86687-86694. doi: 10.18632/oncotarget.13417. PMID: 27893411; PMCID: PMC5349945.

 Abstract

Emerging evidence suggests a role of receptor activator of nuclear factor κB (RANK)/RANK ligand (RANKL) signaling in breast cancer development. Lower osteoprotegerin (OPG) levels, the endogenous decoy receptor for RANKL which competes with RANK for binding of RANKL, has been reported among BRCA mutation carriers. Whether low OPG levels contribute to the high breast cancer risk in this population is unknown. OPG concentrations were measured in plasma of 206 cancer-free BRCA mutation carriers using an enzyme-linked immunosorbent assay. Subjects were categorized as high vs. low based on the median of the entire cohort (95 ng/mL) and followed for a new diagnosis of breast cancer. Cumulative incidence by baseline plasma OPG concentration was estimated using Kaplan-Meier survival analysis. Cox proportional hazards models were used to estimate the adjusted hazard ratios for the association between plasma OPG and breast cancer risk. Over a mean follow-up period of 6.5 years (range 0.1-18.8 years), 18 incident breast cancer cases were observed. After ten years of follow-up, the cumulative incidence of breast cancer among women with low OPG was 21%, compared to 9% among women with high OPG (P-log rank = 0.046). After multivariate adjustment, women with high plasma OPG had a significantly decreased risk of developing breast cancer, compared to women with low OPG (HR = 0.25; 95%CI 0.08-0.78; P = 0.02). These data suggest that low OPG levels are associated with an increased risk of BRCA-associated breast cancer. Targeting RANK signalling may represent a plausible, non-surgical prevention option for BRCA mutation carriers.

Human plasma Osteoprotegerin (OPG) was quantified using the Biomedica OPG ELISA – cat. no. BI-20403 .

Breast Cancer – the most common cancer worldwide

Breast cancer (BC) is the most commonly occurring cancer in women and the most common cancer overall.  Although it is mostly found in women, it can affect men as well. Breast tissue in men can also become malignant. Though male BCs are rare and occur in 1% of all BCs, men are often diagnosed at a more advanced stage. The delay in seeking medical attention often results in late presentation and poor prognosis.

October is breast cancer month  – raising global awareness on risks, the importance of screenings, and the options of treatment.

 

Related links

• Breast cancer statistics
• What’s the difference? Male breast cancer and female breast cancer.

 

Further readings

• Understanding breast cancer as a global health concern.

Wilkinson L et al., Br J Radiol. 2022.  PMID: 34905391; PMCID. Full text

 

Abstract

Breast cancer is now the most commonly diagnosed cancer in the world. The most recent global cancer burden figures estimate that there were 2.26 million incident cases in 2020 and the disease is the leading cause of cancer mortality in women worldwide. The incidence is strongly correlated with human development, with a large rise in cases anticipated in regions of the world that are currently undergoing economic transformation. Survival, however, is far less favourable in less developed regions. There are a multitude of factors behind disparities in the global survival rates, including delays in diagnosis and lack of access to effective treatment. The World Health Organization’s new Global Breast Cancer Initiative was launched this year to address this urgent global health challenge. It aims to improve survival across the world through three pillars: health promotion, timely diagnosis, and comprehensive treatment and supportive care. 

• Global challenges and policy solutions in breast cancer control.

Trapani D et al.,  Cancer Treat Rev. 2022. PMID: 35074727.

 Cancer research

Prognostic exploratory biomarkers

Circulating biomarkers have the potential to provide valuable insight into disease progression. 

 

Discover Biomarker ELISA kits for cancer research – developed and manufactured by Biomedica

•  Neuropilin-1  

regulator of tumor biology – expressed by endothelial cells – isoform-specific receptors for VEGF – checkpoint target –  association with poor prognosis in BC patients.

• Periostin

extracellular matrix protein – novel therapeutic target –  marker of glioma malignancy and potential tumor recurrence – high serum levels associated with a poor survival in BC patients.

• Semaphorin 4D

glycoprotein – emerging clinical biomarker and therapeutic target in cancer – association with cancer progression and the bone metastases.

• Osteoprotegerin, RANKL

contribution to development of bone metastases an –in earlier stages of cancer influence on tumor biology.

• LRG1

Leucine-rich alpha-2-glycoprotein 1  – LRG1 – involved in pathogenic angiogenesis in cancer – wide spread in the microenvironment of numerous tumors – contributes to vascular dysfunction – potential therapeutic target.

• Angiopoietin-2

ANG2 – ANGPT2 – glycoprotein – drives vessel growth – immune target – involved in resistance to anti-VEGF therapy – possible predictive and prognostic biomarker for immune checkpoint therapy.

• IL-6

important cytokine during breast cancer progression – IL6 triggers activation of STAT2 in breast tumors – soluble factor IL6 could be used for early diagnosis of BC or prevent development of metastasis to the bone.  

• VEGF  

Vascular growth factor – potent cytokine that induces tumor angiogenesis – subtype VEGFA and VEGF Receptor -2 are therapeutical targets.

Further information on our products can be found on our website

International agency for research on cancer – WHO

BIOMEDICA & QUALITY

Looking for a reliable ELISA kit for your research?

Use Biomedica kits as your new standard for trustworthy results.

Reliable ELISA kits 

From product development to manufacturing, Biomedica ELISA kits go through a complete validation process.

Contact us for your special evaluation discount by email info@bmgrp.com or call us at our headquarter in Vienna / Austria: +43 1 29107 45 .

We validate our ELISA assays according to international quality standards

The international recognized regulatory forces that issue technical quality guidelines for the validation of bioanalytical methods are:

• FDA (US Food and Drug Administration)
• EMEA (European Medicines Agency)
• ICH (International Conference on Harmonization of Technical Requirements for Registration of Pharmaceuticals for Human Use)

Their main objective is to demonstrate the reliability of a test to determine the concentration of an analyte in a specific biological matrix (e.g. serum, plasma, urine).

Reliable ELISA kits for your research

All Biomedica immunoassays are validated for multiple sample matrices according to above quality guidelines. Our validation experiments include:  

• Accuracy
• Dilution linearity – parallelism
• Specificity
• Cross-reactivity
• Sensitivity
• Precision
• Calibration
• Stability

We Guarantee the Performance of our Products – reliable ELISA kits 

Understanding the importance of high quality, we at Biomedica we offer fully validated ELISA kits for your research. 

To ensure continued customers’ satisfaction, we strive to meet the highest quality standards and continuously work on improving both our products and manufacturing process.

For further details on our validation process come and visit our quality site.

Test our high-quality ELISA kits for your research and ask for your special evaluation discount.

Contact us by email  info@bmgrp.com or call us at our office in Vienna – Austria: +43 1 29107 45 .

Cancer research for the prevention and early recognition of cancer

Promoting cancer research to beat cancer: prevention and early detection of cancer is essential to control the disease. Understanding the causes of cancer and the ability to detect cancer sooner has a great impact on the survival and the outcome of patients.

Subsequently, early detection strategies and progress in diagnostic procedures will help to develop treatments to control the disease.

Promoting cancer research:

Exploring new biomarkers may help to identify the disease early.

Biomedica offers a range of novel biomarker ELISA kits for your cancer research.

Novel research biomarkers for cancer research include:  Periostin,  Neuropilin-1,  Semaphorin 4D

A range of cytokine ELISA kits complete  the panel:  Interleukin-6,  VEGF,  Angiopoietin-2 .

See our comprehensive “Oncology Biomarker Brochure “ for more information or contact us directly info@bmgrp.com .

We will be glad to meet you.

RELATED LITERATURE:

Promoting Cancer Early Diagnosis –  https://www.who.int

“Early diagnosis of cancer focuses on detecting symptomatic patients as early as possible so they have the best chance for successful treatment. When cancer care is delayed or inaccessible there is a lower chance of survival, greater problems associated with treatment and higher costs of care. Early diagnosis improves cancer outcomes by providing care at the earliest possible stage and is therefore an important public health strategy in all settings…”   Read more

Understanding mechanisms of cancer initiation and development supports the need for an implementation of primary and secondary cancer prevention.

Pelosi E, Castelli G, Testa U.Ann Ist Super Sanita. 2019 Oct-Dec;55(4):371-379. doi: 10.4415/ANN_19_04_11. PMID: 31850865.

Abstract

The burden of cancer is increasing worldwide, with a continuous rise of the annual total cases. Although mortality rates due to cancer are declining in developed countries, the total number of cancer deaths continues to rise due to the increase in the number of aged people. Three main causes of cancer have been described, represented by environmental factors, hereditary factors and random factors related to defects originated during cell replication. The frequency of cancers is very different for the various tissues and there is great debate on the extent of the specific contribution of environmental factors and random factors (due to “bad luck”) to cancer development. However, there is consensus that about 50% of all cases of cancer are related to environment and are preventable. Although a part of cancers is related to intrinsic mechanisms non preventable of genetic instability, it is evident that implementation of primary and secondary prevention measures is the only affordable strategy to meet from a medical and economic point of view the tremendous pressure created on healthcare structures by the increased cancer burden. It is time to bypass the paradox of disease prevention: celebrated in principle, resisted in practice.

Ovarian Cancer Biomarkers: Moving Forward in Early Detection

Bonifácio VDB. Adv Exp Med Biol. 2020;1219:355-363. doi: 10.1007/978-3-030-34025-4_18. PMID: 32130708.

Abstract

Ovarian cancer is a silent cancer which rate survival mainly relays in early stage detection. The discovery of reliable ovarian cancer biomarkers plays a crucial role in the disease management and strongly impact in patient’s prognosis and survival. Although having many limitations CA125 is a classical ovarian cancer biomarker, but current research using proteomic or metabolomic methodologies struggles to find alternative biomarkers, using non-invasive our relatively non-invasive sources such as urine, serum, plasma, tissue, ascites or exosomes. Metabolism and metabolites are key players in cancer biology and its importance in biomarkers discovery cannot be neglected. In this chapter we overview the state of art and the challenges facing the use and discovery of biomarkers and focus on ovarian cancer early detection.

 Keywords: Cancer biomarkers; Early detection; Metabolomics; Ovarian cancer; Proteomics; Urine biomarkers.

 

Back to school – sports training has positive outcomes

Strength training for children and adolescents enhances bone health

Strength training for children and adolescents is becoming more important as part of sport training and after-school fitness programs. Consequently, health problems due to inactivity, sedentary lifestyle and being overweight have resulted in increased interest in strength and resistance training (1). Today there is ample evidence that youth resistance training is safe and effective and improves motor skills, reduces fat mass, and enhances bone health. In addition, various performance markers such as muscle strength, power and overall health also improve (2-5). In adults, weight-bearing impact exercise such as jumping or hopping in addition to strength training can improve bone health. Among these, resistance training is the most promising intervention to maintain or increase bone mass and density (5, 6). 

Subsequently, measuring serum bone related biomarkers can be helpful in understanding normal and pathological processes that reflect bone cell activities in the skeleton.

Check out Biomedica´s high quality bone marker ELISA kits: Sclerostin,  RANKL, OPG,  DKK-1,  FGF23 (C-terminal), FGF23 intact and other 

RELIABLE MANUFACTURING ENSURES CONSISTENT RESULTS

Human Sclerostin ELISA kit 

√  The internationally most referenced Sclerostin ELISA!
√  Extensively validated according to international quality guidelines
√  Only 20µl sample / well – low sample volume

References on strength training for children and adoloscents 

1. Resistance Training for Children and Adolescents.
   Stricker PR, Faigenbaum AD, McCambridge TM; COUNCIL ON SPORTS MEDICINE AND FITNESS Pediatrics. 2020. 145(6):e20201011. doi: 10.1542/peds.2020-1011.
2. Performance – and health-related benefits of youth resistance training – Leistungs- und gesundheitsbezogene Wirkungen von Krafttraining mit Heranwachsenden.
   H.Chaabene H et al., 2020. Sports Orthopaedics and Traumatology; 36: 231-240.
3. Strength training for children and adolescents: benefits and risks.
   Barbieri D, Zaccagni L. Coll Antropol. 2013. 37 Suppl 2:219-25. PMID: 23914510.
4. Strength training for children and adolescents.
   Faigenbaum A. 2000. Clinics in Sports Medicine; 593-619.
5. Exercise and Sports Science Australia (ESSA) position statement on exercise prescription for the prevention and management of osteoporosis.
   Beck BR, Daly RM, Singh MA, Taaffe DR. J Sci Med Sport. 2017;20:438–445. 
6. Effects of Resistance Exercise on Bone Health.
   Hong AR, Kim SW. Endocrinol Metab (Seoul). 2018. 33(4):435-444. doi: 10.3803/EnM.2018.33.4.435. PMID: 30513557; PMCID: PMC6279907.

 

RELATED LITERATURE

Youth Resistance Training: The Good, the Bad, and the Ugly-The Year That Was 2017.
Faigenbaum AD. Pediatr Exerc Sci. 2018. 1;30(1):19-24. doi: 10.1123/pes.2017-0290. PMID: 29424264.

Abstract

The good news is that a growing body of evidence recognizes resistance training as foundational to long-term physical development. Original research and reviews published in 2017 conclude that early exposure to developmentally appropriate resistance training can improve markers of health, increase muscular fitness, enhance physical literacy, and reduce the risk of injury in young athletes. Although the papers discussed in the commentary add to our understanding of the pleiotropic benefits of youth resistance training, they also raise concerns. As measures of muscular strength and power have been found to track from childhood to adulthood, the bad news is that youth with low levels of muscular fitness tend to become weak adults who are at increased risk for functional limitations and adverse health outcomes. Furthermore, global participation in youth resistance training is falling far short of public health recommendations, and these ugly trends will likely impact the health and well-being of future generations. A change in current attitudes and common practices is urgently needed to educate parents, practitioners, and clinicians about the potential benefits of resistance training for all children and adolescents, not only young athletes.

Metabolic syndrome: Operational definitions and aerobic and resistance training benefits on physical and metabolic health in children and adolescents.

Leister KR, Cilhoroz BT, Rosenberg J, Brown EC, Kim JY Diabetes Metab Syndr. 2022. 16(6):102530. doi: 10.1016/j.dsx.2022.102530. PMID: 35709585.

Watch this on how kids can get stronger muscles:   “Top 10 Kids Exercises To Get Stronger Muscles”

Strength training for children and adolescents enhances bone health