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• NT-proBNP in end-stage kidney disease

Cardiovascular disease is the leading cause of death in patients with end-stage renal disease on dialysis (1). In an attempt to understand the underlying mechanisms of the high mortality among dialysis patients, other than the traditional cardiovascular risk factors, researchers investigated the association between gut permeability, circulating bacterial fragments, and volume overload in peritoneal dialysis (PD) patients (2).

Gut permeability, circulating bacterial fragments and measures of congestion in peritoneal dialysis.

In this prospective observational study, 108 consecutive adult incident PD patients were recruited and circulating bacterial fragments, N-terminal pro B-type natriuretic peptide (NT-proBNP), calprotectin and zonulin levels were evaluated (2).

NT-proBNP in end-stage kidney disease

Markers of fluid overload

NT-proBNP is recognized as a marker of marker of left ventricular strain and intravascular fluid overload (3, 4).

Fluid overload was quantified by serum levels of NT-proBNP with the Biomedica ELISA kit (Biomedica-cat. no. SK-1204) and with multi-frequency bioimpedance spectroscopy (2).

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

√  CE-marked – for IVD use in the EU
√  EASY – simple 2 step protocol, can be run in every lab
√  High and low kit controls included 
√  RELIABLE – validated according to international quality guidelines (see validation data )
√  TRUSTED – widely cited in 125 publications

 

Example of a Biomedica ELISA Assay Kit

 

Literature                                                                    

1.   Cardiovascular complications in chronic kidney disease: a review from the European Renal and Cardiovascular Medicine Working Group of the European Renal Association. Zoccali C, Mallamaci F, Adamczak M, de Oliveira RB, Massy ZA, Sarafidis P, Agarwal R, Mark PB, Kotanko P, Ferro CJ, Wanner C, Burnier M, Vanholder R, Wiecek A. Cardiovasc Res. 2023 Sep 5;119(11):2017-2032. doi: 10.1093/cvr/cvad083. PMID: 37249051; PMCID: PMC10478756.

2.   Gut permeability, circulating bacterial fragments and measures of congestion in peritoneal dialysis. Li C, Ng JK, Chan GC, Fung WW, Lai KB, Poon PY, Luk CC, Chow KM, Szeto CC. Clin Kidney J. 2024 Mar 6;17(3):sfae056. doi: 10.1093/ckj/sfae056. PMID: 38516523; PMCID: PMC10956420. 

3.   NT-proBNP, fluid volume overload and dialysis modality are independent predictors of mortality in ESRD patients. Paniagua R, Ventura MD, Avila-Díaz M, Hinojosa-Heredia H, Méndez-Durán A, Cueto-Manzano A, Cisneros A, Ramos A, Madonia-Juseino C, Belio-Caro F, García-Contreras F, Trinidad-Ramos P, Vázquez R, Ilabaca B, Alcántara G, Amato D. Nephrol Dial Transplant. 2010 Feb;25(2):551-7. doi: 10.1093/ndt/gfp395. Epub 2009 Aug 12. PMID: 19679559.

4.   Determination of volume overload by bioelectrical impedance analysis and NT-proBNP in diabetic pre-dialysis patients. Acta Endocrinol (Buchar). Yildirim Y, Kara AV, Kilinç F, Aydin F, Aydin E, Yilmaz Z, Kadiroglu AK, Yilmaz ME. 2016 Jan-Mar;12(1):19-25. doi: 10.4183/aeb.2016.19. PMID: 31258795; PMCID: PMC6586753.

Proficiency Testing is an external validation of diagnostic methods performed by accredited laboratories (1) as a measure to comply with international quality standards. Through participation in inter-laboratory comparisons such as proficiency testing programs, customers using a diagnostic method (e.g. an ELISA assay), can be assured that results generated from the assay comply with internationally set quality standards.

NT-proBNP assay successfully passes cardiac marker survey

NT-proBNP as a marker of heart failure

Heart failure is a significant cause of morbidity and mortality worldwide. Circulating biomarkers reflecting disease-related pathways that are involved in the development and progression of heart failure (HF) can be helpful in assisting  clinicians in the early diagnosis and management of HF patients.

NT-proBNP (N-terminal pro B-type Natriuretic Peptide) is a cardiac protein fragment that is secreted into the bloodstream when the heart muscle stretches (under stress or damage). NT-proBNP is recognized as the hallmark biomarker for heart failure diagnosis and prognosis (2, 3).

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

• CE-marked – for IVD use in the EU
• Proficiency tested – click here to access the certificate 
• Flexible – can be run in every lab
• Accurate – two controls included
• Easy – 2 step protocol
• Trusted – widely cited in 125 publications

 

Example of a Biomedica ELISA Assay kit

Developed & manufactured by Biomedica 

 

Literature

1. RfB- reference institute for bioanalytics– proficiency testing
2. NT-proBNP: The Gold Standard Biomarker in Heart Failure. PM McKie PM and JC Jr Burnett. J Am Coll Cardiol. 2016; Dec 6;68(22):2437-2439. doi: 10.1016/j.jacc.2016.10.001. PMID: 27908348.
3. Biomarkers for the diagnosis and management of heart failure. Castiglione V, Aimo A, Vergaro G, Saccaro L, Passino C, Emdin M.Heart Fail Rev. 2022 Mar;27(2):625-643. doi: 10.1007/s10741-021-10105-w. Epub 2021 Apr 14. PMID: 33852110; PMCID: PMC8898236.

Smoking is a major risk factor for a wide range of disorders, including cardiovascular, respiratory diseases, and cancers. Cigarette smoke generates a high level of reactive oxygen species (ROS), causing oxidative stress and cellular damage (1-3). Among the ROS, aldehyds are the main compounds that are implicated in the process of tobacco-induced diseases (3).

Autoantibodies against oxidized-LDL in smokers

Smoking is associated with increased levels of Autoantibodies against oxidized-LDL

In smokers, low density lipoprotein (LDL) is more prone to oxidation due to an excessive presence of reactive ROS, leading to elevated levels of antibodies against oxidized LDL (anti-oxLDL Ab). In healthy individuals no significant differences in anti-oxLDL Ab levels have been observed between smokers and non-smokers. However, moderately elevated levels of anti-oxLDL Ab have been noted in hypercholesterolemic patients (4).

In the same patient group researchers found that foods containing olive oil and lycopene can significantly contribute to reducing LDL-induced oxidative stress (5). The authors suggest that lycopene-olive oil can be used as a supplement to reduce oxidative stress and the inflammatory process in hypercholesterolemic subjects, thereby potentially lowering the risk of atherosclerosis (5).

Function of Low-density lipoprotein (LDL)

Low-density lipoprotein (LDL) is responsible for transporting cholesterol from the liver to various tissues and cells in the body. It is used for essential functions like cell repair, cell membrane structure  and hormone production. where it is used fo and other lipids through the bloodstream (6).

Smoking leads to a rise of LDL cholesterol levels, which contributes to plaque buildup in arteries, increasing heart disease and the risk of stroke. Smoking promotes the oxidation of LDL particles, and ox LDL is more likely to adhere to artery walls, leading to atherosclerosis (7).

Antibodies against oxidized LDL (anti-oxLDL Ab).

Oxidized low-densitity lipoprotein (oxLDL) plays an important role in in the development of atherosclerosis. Autoantibodies against oxidatively modified LDL particles can serve as a parameter that consistently reflects the ongoing oxidation processes taking place in vivo.  Studies demonstrate increased levels of autoantibodies against oxLDL in the bloodstream of individuals with coronary artery disease (4)

Several studies reported a significant association between anti-oxLDL levels and the subsequent development of Atherosclerosis-related cardiovascular outcomes (4, 8, 9).

 Measurement of  Antibodies against oxidized LDL (anti-oxLDL Ab)

Autoantibodies targeting oxidatively modified LDL particles can be measured serum with an ELISA assay

Anti-Oxidized LDL Autoantibody ELISA (oLAB) Assay (cat. no. BI-20032)

Features

• Widely cited
• Results in 2,5 hours
• 2 controls included

 

 

Literature

1. Cigarette Smoke-Induced Reactive Oxygen Species Formation: A Concise Review. Antioxidants (Basel). Seo YS, Park JM, Kim JH, Lee MY. 2023 Sep 7;12(9):1732. doi: 10.3390/antiox12091732. PMID: 37760035; PMCID: PMC10525535.
2. Relationships among smoking, oxidative stress, inflammation, macromolecular damage, and cancer. Caliri AW, Tommasi S, Besaratinia A. Mutat Res. 2021 Jan-Jun;787:108365. doi: 10.1016/j.mrrev.2021.108365. Epub 2021 Jan 11. PMID: 34083039; PMCID: PMC8287787.
3. Exocyclic DNA adducts and oxidative stress parameters: useful tools for biomonitoring exposure to aldehydes in smokers. Alamil H, Colsoul ML, Heutte N, Van Der Schueren M, Galanti L, Lechevrel M. Biomarkers. 2024 Apr 2:1-7. doi: 10.1080/1354750X.2024.2333361. Epub ahead of print. PMID: 38506499.
4. Cigarette smoking potentiates endothelial dysfunction of forearm resistance vessels in patients with hypercholesterolemia. Role of oxidized LDL. Heitzer T, Ylä-Herttuala S, Luoma J, Kurz S, Münzel T, Just H, Olschewski M, Drexler H. Circulation. 1996 Apr 1;93(7):1346-53. doi: 10.1161/01.cir.93.7.1346. PMID: 8641023.
5. Beneficial Effects of Olive Oil Enriched with Lycopene on the Plasma Antioxidant and Anti-Inflammatory Profile of Hypercholesterolemic Patients. Martínez Álvarez JR, Lopez Jaen AB, Cavia-Saiz M, Muñiz P, Valls-Belles V. Antioxidants (Basel). 2023 Jul 20;12(7):1458. doi: 10.3390/antiox12071458. PMID: 37507996; PMCID: PMC10376681.
6. Biochemistry, Low Density Lipoprotein, Senthil K. Venugopal; McDamian Anoruo; Ishwarlal Jialal. 2024, StatPearls Publishing LLC.
7. Smoking and small, dense low-density lipoprotein particles: cross-sectional study. Urahama N, Iguchi G, Shimizu M, Fujihira K, Kobayashi S, Baba H. Nicotine Tob Res. 2008 Aug;10(8):1391-5. doi: 10.1080/14622200802238852. PMID: 18686187.
8. Oxidized LDL to autoantibodies against oxLDL ratio – The new biomarker associated with carotid atherosclerosis and cardiovascular complications in dialyzed patients. Pawlak, K., Mysliwiec, M., Pawlak, D., 2012. Atherosclerosis 224, 252–257.
9. Circulating oxidized low density lipoprotein, autoantibodies against them and homocysteine serum levels in diagnosis and estimation of severity of coronary artery disease. Faviou, E., Vourli, G., Nounopoulos, C., Zachari, A., Dionyssiou-Asteriou, A., 2005. Free Radic. Res. 39, 419–429.

Atherosclerosis is a chronic inflammatory disease of the arterial wall leading to the formation of atherosclerotic plaques (1). It is the primary cause of cardiovascular disease, affecting millions of individuals every year. Despite advances in our understanding of the disease, the exact mechanisms involved in plaque formation are not yet fully understood.  

LRG-1 promotes calcification in atherosclerosis

In an attempt to gain insight into the complex processes in the development of atherosclerosis, researchers identified for the first time the molecule Leucine-Rich Alpha-2 Glycoprotein-1 (LRG1) that contributes directly to vascular calcification in mice (2). The authors suggest that LRG1 is linked to the development of plaque complications in patients with atherosclerosis. LRG1 may be a novel therapeutic target to slow down the calcification process that remains a challenge in patients with diabetes and chronic renal failure (2). Learn more about the study: Leucine-Rich Alpha-2 Glycoprotein 1 Accumulates in Complicated Atherosclerosis and Promotes Calcification. 

About Leucine-Rich α-2 Glycoprotein 1 (LRG1)

LRG1* is a protein that is primarily synthesized and secreted by the liver and immune cells. It is involved in numerous conditions including lung, kidney, and heart disease. The pathogenic roles of LRG1 in these diseases are often linked to its effects on the vasculature (3). A recent review by Dritsoula A. and colleauges  summarizes the multifaceted role of LRG in disrupting the vasculature. LRG has been reported to damage blood vessels in conditions such as cancer, diabetes, chronic kidney disease, ocular disease, and lung disease. Furthermore, therapeutic targeting of LRG1 has been widely proposed as a strategy to restore quienscent endothelium and normalize vasculature (4).

*LRG also named LRG1 (leucine-rich alpha-2-glycoprotein) is a glycoprotein with a molecular mass of 38.2 kDa (https://www.uniprot.org/uniprot/P02750). It is encoded by the human gene LRG-1.  

 

Quantification of LRG1 in human samples

LRG1 can easily be quantified in human samples (serum, plasma, urine, cell culture supernatants) with a conventional ELISA* assay method.

*The ELISA technique is an immunoassay method that provides a tool to detect or quantify the concentration of a specific analyte in a sample (serum, plasma, urine, cell-culture supernatant).

Check out the LRG1 ELISA protocol booklet – day test and our poster on “Novel ELISA for the quantification of human leucine-rich α-2 glycoprotein (LRG) in serum and plasma”

 

LRG ELISA – developed and manufactured by BIOMEDICA (cat. no. BI-LRG)

• Full validation – data can be found here.
• LRG values available for normal and pathological samples
• Results in 3.5 hours
• ELISA kit includes 2x controls, 7x standards (for ready standard curve)

 

Contact us for your special study discount! 

 

Literature

1. The changing landscape of atherosclerosis. Libby P. Nature. 2021 Apr;592(7855):524-533.  PMID: 33883728.
2. Leucine-Rich Alpha-2 Glycoprotein 1 Accumulates in Complicated Atherosclerosis and Promotes Calcification. Grzesiak L, Amaya-Garrido A, Feuillet G, Malet N, Swiader A, Sarthou MK, Wahart A, Ramel D, Gayral S, Schanstra JP, Klein J, Laffargue M. Int J Mol Sci. 2023 Nov 20;24(22):16537.  PMID: 38003727.
3. LRG1: an emerging player in disease pathogenesis. Camilli C, Hoeh AE, De Rossi G, Moss SE, Greenwood J. J Biomed Sci. 2022 Jan 21;29(1):6.  PMID: 35062948.
4. The disruptive role of LRG1 on the vasculature and perivascular microenvironment. Dritsoula A, Camilli C, Moss SE, Greenwood J.Front Cardiovasc Med. 2024 Apr 30;11:1386177. PMID: 38745756.

 

 

 

Rheumatoid arthritis is a chronic inflammatory disorder that affects not only the joints but can also damage other parts of the body including the skin, lungs (1), heart and blood vessels. Patients with RA have a notable 50-70% increased risk of developing cardiovascular diseases compared to the general population (2).

Effects of Tofacitinib Therapy on Angiogenic Biomarkers in Rheumatoid Arthritis

In a recent study, highlighting the Biomedica NT-proBNP ELISA Kit*, researchers investigated for the first time the effects of 1-year Tofacitinib therapy on angiogenic biomarkers in relation to vascular inflammation and function as well as clinical markers in patients with rheumatoid arthritis (RA) (3). Tofacitinib is a medication used to treat RA. It inhibits Janus kinase enzymes which are involved in cytokine signaling leading to inflammation and symptoms of RA (4).

*Serum NT-proBNP (pmol/l) concentrations were detected by commercially available ELISA kits (NT-proBNP ELISA, Biomedica, Vienna, Austria)

NT-proBNP ELISA Kit (cat. no. SK-1204)

• Quality – CE marked – for IVD use in the EU (and proficiency tested*)
• Easy – simple protocol, kit includes two controls
• Reliable – fully validated 
• Trusted – Cited in 125 publications

Download the Biomedica NT-proBNP ELISA Assay Proficiency Testing Certificate here

 

 

About Proficiency Testing

Proficiency Testing offers a report enabling laboratories to benchmark their data against data from other laboratories globally. Conducted within a ring trial program, this testing is executed by accredited laboratories specialized on proficiency testing.  Proficiency testing oversees the performance of individual laboratories for specific tests like cardiac biomarker assays.

 

Effects of 1-year tofacitinib therapy on angiogenic biomarkers in rheumatoid arthritis. Kerekes G et al., Rheumatology (Oxford). 2023

Abstract 

Objectives – Cardiovascular (CV) morbidity and mortality, and perpetuated synovial angiogenesis have been associated with RA. In our study we evaluated angiogenic factors in relation to vascular inflammation and function, and clinical markers in RA patients undergoing 1-year tofacitinib therapy.

Methods – Thirty RA patients treated with either 5 mg or 10 mg twice daily tofacitinib were included in a 12-month follow-up study. Eventually, 26 patients completed the study and were included in data analysis. Levels of various angiogenic cytokines (TNF-α, IL-6), growth factors [VEGF, basic fibroblast (bFGF), epidermal (EGF), placental (PlGF)], cathepsin K (CathK), CXC chemokine ligand 8 (CXCL8), galectin-3 (Gal-3) and N-terminal prohormone brain natriuretic peptide (NT-proBNP) were determined at baseline, and at 6 and 12 months after initiating tofacitinib treatment. In order to assess flow-mediated vasodilation, common carotid intima-media thickness (ccIMT) and carotid-femoral pulse-wave velocity, ultrasonography was performed. Synovial and aortic inflammation was also assessed by ¹⁸F-fluorodeoxyglucose-PET/CT.

Results – One-year tofacitinib therapy significantly decreased IL-6, VEGF, bFGF, EGF, PlGF and CathK, while it increased Gal-3 production (P < 0.05). bFGF, PlGF and NT-proBNP levels were higher, while platelet-endothelial cell adhesion molecule 1 (PECAM-1) levels were lower in RF-seropositive patients (P < 0.05). TNF-α, bFGF and PlGF correlated with post-treatment synovial inflammation, while aortic inflammation was rather dependent on IL-6 and PECAM-1 as determined by PET/CT (P < 0.05). In the correlation analyses, NT-proBNP, CXCL8 and Cath variables correlated with ccIMT (P < 0.05).

Conclusions – Decreasing production of bFGF, PlGF or IL-6 by 1-year tofacitinib therapy potentially inhibits synovial and aortic inflammation. Although NT-proBNP, CXCL8 and CathK were associated with ccIMT, their role in RA-associated atherosclerosis needs to be further evaluated.

 

Literature

1. Identification, Monitoring, and Management of Rheumatoid Arthritis-Associated Interstitial Lung Disease. Koduri G and  Solomon JJ. Arthritis Rheumatol, 2023; Dec;75(12):2067-2077. doi: 10.1002/art.42640.
2. Association of cardiovascular risks in rheumatoid arthritis patients: Management, treatment and future perspectives. Nishant Johri et al., 2023; Health Sciences Review, Vol 8.
3. Effects of 1-year tofacitinib therapy on angiogenic biomarkers in rheumatoid arthritis. Kerekes G et al., Rheumatology (Oxford), 2023; 62(SI3):SI304-SI312. doi: 10.1093/rheumatology/kead502. 
4. Efficacy and safety of JAK inhibitors in rheumatoid arthritis: update for the practising clinician. Szekanecz Z et al.,  Nat Rev Rheumatol, 2024; 20(2):101-115. doi: 10.1038/s41584-023-01062-9.

 

 

 

 

Lyme disease (LD) or Lyme Borreliosis is a bacterial infection that can be transmitted to humans by infected ticks. It is estimated that about 700 000 individuals in Europe and in the United States are infected with LD every year, though the number of unrecorded or misdiagnosed cases could be higher (1).

Symptoms include fever, headache, joint pain, and swollen lymph nodes. Lyme disease is often accompanied by a distinctive expanding rash (Erythema Migrans) which starts at the site of the tick bite (2).  In most cases when LD is diagnosed early, treatment with antibiotics is very effective. However, without treatment, the infection can spread to different areas of the body, impacting the joints, heart, and nervous system (3). Early diagnosis and treatment are essential for managing Lyme disease effectively.

The Biomedica Borrelia ELISA Assays were utilized in a recent study to estimate the incidence of symtopmatic Lyme Borreliosis cases in Lubin, Poland in 2021:  Estimated Incidence of Symptomatic Lyme Borreliosis Cases in Lublin, Poland in 2021. Colby E et al., Microorganisms. 2023; 11(10):2481. The authors concluded that “after adjusting for under-ascertainment, the estimated number of symptomatic LB cases in Lublin in 2021 was 6204 (population-based incidence: 467.6/100,000). After adjustment for under-ascertainment, the incidence of symptomatic LB in Lublin, Poland, is high.”

Serological Testing for Lyme Disease with BIOMEDICA ELISA Kits

The Borrelia assays from Biomedica use recombinant antigens to specifically detect IgM and IgG antibodies against the immunodominant antigens of the three genospecies (Borrelia burgdorferi sensu stricto, Borrelia afzelii and Borrelia garinii) and thus offer:

• High sensitivity and specificity that is confirmed by clinical samples
• Standardized CSF and serum analysis available
• For manual and automated testing
• No extra RF stripping necessary
• Widely cited
• CE marked – for IVD use in the EU

 

Borrelia IgM ELISA (cat. no. BI-21042)

ENZYME IMMUNOASSAY FOR THE QUALITATIVE OR QUANTITATIVE DETERMINATION OF IgG ANTIBODIES TO BORRELIA IN PLASMA, SERUM OR CEREBROSPINAL FLUID

The following recombinant antigens are utilized in the Biomedica Borrelia IgM ELISA Assay:

• p21  OspC – B. afzellii (pKo)
• p21  OspC – B. garinii (20047)
• p41/I – B. bavariensis (pBi)
• VIsE – fusion proteins of different Borrelia genospecies 

 

Borrelia IgG ELISA (cat. no. BI- 21032)

ENZYME IMMUNOASSAY FOR THE QUALITATIVE OR QUANTITATIVE DETERMINATION OF IgG ANTIBODIES TO BORRELIA IN PLASMA, SERUM OR CEREBROSPINAL FLUID

The following recombinant antigens are utilized in the Biomedica Borrelia IgG ELISA Assay:

• p21 – OspC – B. burgdorferi sensu stricto (B31), B. garinii (20047)
• p18 – B. afzelii (pKo)
• p100 – B. afzelii (pKo)
• VIsE – fusion protein of different Borrelia genospecies

 

Literature

1. Lyme borreliosis diagnosis: state of the art of improvements and innovations. Guérin M et al., 2023; BMC Microbiol, 23(1):204.
2. Lyme borreliosis. Steere AC et al., 2016 ; Rev Dis Primers. 2:16090.
3. Neurologic manifestations of Lyme Borreliosis. Summer G et al., 2019; Rev Neurol (Paris). 175(7-8):417-419.

 

 

The Biomedica soluble SEMAPHORIN 4D Assay Kit (sSEMA4D) was utilized in a recent study assessing the role of sSEMA4D in critically ill and septic patients as a biomarker for prediction and disease severity or survival (1). The results revealed that blood SEMA4D concentrations are increased in patients with liver cirrhosis and are positively correlated with liver function tests. The study suggests that sSEMA4D may be associated with hepatic injury and inflammation and may serve as a potential biomarker in critically ill patients with liver cirrhosis.

Semaphorin 4D in critically ill patients

About Semaphorin 4D

Semaphorin 4D (Sema4D) also known as CD100 is a member of the Sema4D family with established immunregulatory functions (2). Sema 4D exists as a membrane bound and in a soluble form. It is expressed on immune cells and is upregulated upon cellular activation resulting in shedding of its extracellular soluble Sema4D domain. This soluble biologically active form has a molecular weight of 120KD and can be measured in blood samples (3, 4). Due to its regulatory role in the immune system the potential application of Sema4D as a diagnostic marker and as a therapeutic target for the treatment of immunological disorders is currently under investigation (5).

Biomedica soluble SEMAPHORIN 4D (sSEMA4D) ELISA Assay Kit (cat. no. BI-20405)

Features & Benefits

• Only Sema4D ELISA assay that is fully validated
• 10 µl / well sample volume – no sample predilution
• Reference values provided

Links to soluble Semaphorin 4D ELISA  – protocol booklet  –  validation data file

 

 

Literature 

1. Soluble Semaphorin 4D Serum Concentrations Are Elevated in Critically Ill Patients with Liver Cirrhosis and Correlate with Aminotransferases. Abu Jhaisha S, Hohlstein P, Yagmur E, Köller V, Pollmanns MR, Adams JK, Wirtz TH, Brozat JF, Bündgens L, Hamesch K, Weiskirchen R, Tacke F, Trautwein C, Koch A. Diagnostics (Basel). 2024 Feb 8;14(4):370. doi: 10.3390/diagnostics14040370. PMID: 38396409; PMCID: PMC10887520.
2. The Role of Semaphorin 4D in Bone Remodeling and Cancer Metastasis. Lontos K, Adamik J, Tsagianni A, Galson DL, Chirgwin JM, Suvannasankha A. Front Endocrinol (Lausanne). 2018 Jun 19;9:322. doi: 10.3389/fendo.2018.00322. PMID: 29971044; PMCID: PMC6018527.
3. Soluble SEMA4D/CD100: A novel immunoregulator in infectious and inflammatory diseases. Maleki KT, Cornillet M, Björkström NK. Clin Immunol. 2016 Feb;163:52-9. doi: 10.1016/j.clim.2015.12.012. Epub 2015 Dec 28. PMID: 26732857.
4. A high-sensitivity enzyme immunoassay for the quantification of soluble human semaphorin 4D in plasma. Laber A, Gadermaier E, Wallwitz J, Berg G, Himmler G. Anal Biochem. 2019 Jun 1;574:15-22. doi: 10.1016/j.ab.2019.03.004. Epub 2019 Mar 14. PMID: 30879960.
5. The role of Sema4D/CD100 as a therapeutic target for tumor microenvironments and for autoimmune, neuroimmune and bone diseases. Wu M, Li J, Gao Q, Ye F. Expert Opin Ther Targets. 2016 Jul;20(7):885-901. doi: 10.1517/14728222.2016.1139083. Epub 2016 Jan 28. PMID: 26732941.

 

 

 

Irritable bowel syndrome (IBS) is a common gastrointestinal disorder affecting approximately around 1 in 10 people globally (1).

IBS is characterized by recurrent abdominal pain and changes in bowel habits. The exact causes of IBS is unknown but stress, gastrointestinal infections and a family history of IBS may play a role (1,2). IBS is usually a lifelong problem and symptoms have an effect on daily life. Managing symptoms through diet and lifestyle changes can improve the quality of life  (2, 3, 4) .

Accepted biomarkers for IBS are still missing. Recent studies have shown that Leucine-rich alpha glycoprotein (LRG) may be a novel marker to evaluate disease activity and mucosal healing in IBS (5, 6). Moreover, researchers have recently proposed that serum LRG may be a potential diagnostic marker for pediatric IBD (7).

Biomarkers for Irritable Bowel Syndrome

Leucine-rich alpha glycoprotein (LRG)

Leucine-rich alpha-2 glycoprotein (LRG) (also named LRG1) has a molecular weight of 50 kDa glycoprotein and contains repetitive sequences with a leucine-rich motif. LRG derives predominantly from immune cells i.e. neutrophils, macrophags but is also secreted by intestinal epithelial cells and hepatocytes (8) in response to various cytokines (TNF-alpha, IL-6 and IL-22).

LRG has been identified as an inflammatory biomarker for various immune-mediated diseases including IBD, rheumatoid arthritis, juvenile idiopathic arthritis and others (8).

LRG can be measured in human serum and plasma samples by ELISA Assay

LRG ELISA (cat. no. BI-LRG) – developed and manufactured by BIOMEDICA

• RELIABLE – validated following international quality guidelines (FDA, EMA, ICH) . The validation data can be found here.
• REFERENCE values provided for normal and pathological samples
• OPTIMIZED – assay range optimized for clinical samples, no additional testing required
• EASY – results in 3.5 h (protocol booklet)
• CONVENIENT – all reagents included; 7 pre-diluted standards/calibrators, 2 controls and sufficient assay buffer

 

Example of a Biomedica ELISA Assay Kit

 

Literature

1. Global burden of irritable bowel syndrome: trends, predictions and risk factors. Black CJ et al., Nat Rev Gastroenterol Hepatol. 2020;  17(8):473-486. PMID: 32296140.
2. Irritable bowel syndrome. Ford AC et al., Lancet. 2020; 396(10263):1675-1688. PMID: 33049223.
3. A Discussion of Whether Various Lifestyle Changes can Alleviate the Symptoms of Irritable Bowel Syndrome. Okawa Y. Healthcare (Basel). 2022; 10(10):2011. PMID: 36292457.
4. Irritable bowel syndrome: treatment based on pathophysiology and biomarkers. Camilleri M, Boeckxstaens G. Gut. 2023;72(3):590-599. PMID: 36307180.
5. Leucine-rich alpha-2 glycoprotein as a marker of mucosal healing in inflammatory bowel disease. Yasutomi E et al., Sci Rep. 2021; 27;11(1):11086.
6. Leucine-Rich Alpha-2 Glycoprotein Is a Reliable Serum Biomarker for Evaluating Clinical and Endoscopic Disease Activity in Inflammatory Bowel Disease. Shimoyama Tet al., Inflamm Bowel Dis. 2023; 29(9):1399-1408. 
7. Significance of Serum Leucine-rich Alpha-2 Glycoprotein as a Diagnostic Marker in Pediatric Inflammatory Bowel Disease. Yoshimura S et al., Kobe J Med Sci. 2024; 69(4):E122-E128. PMID: 38379274.
8. Evaluation of Serum Leucine-Rich Alpha-2 Glycoprotein as a New Inflammatory Biomarker of Inflammatory Bowel Disease. Yoshimura Tet a.,  2021; 2021:8825374. PMID: 33623482.

 

 

Traumatic brain injury (TBI) is a sudden injury that causes damages to the brain. It is one of the most common causes of disability and death in adults (1). There are currently no early biomarkers for prognosis in routine clinical use (2).  Apart from the initial injury, victims with TBI face the possibility of secondary neurological damage. Studies have identified TBI as a risk factor for all-cause dementia and Parkinson’s disease (3).

Interleukin-6 in Traumatic Brain Injury

Interleukin (IL)-6 is a proinflammatory cytokine that plays a key role in the immune response to acute neurological injury (1, 4) . Studies have shown that IL-6 may serve as a prognostic biomarker of clinical outcomes after traumatic brain injury (2).

IL-6 can reliably be measured with the highly sensitive IL-6 ELISA assay from Biomedica

IL-6 ELISA (Biomedica, cat. no. BI-IL6)

• HIGHLY SENSITIVE- measurable values in serum and plasma samples
• EASY – ready to use calibrators and controls
• RELIABLE – full validation package

 

 

Literature

1. Interleukin-6 in Traumatic Brain Injury: A Janus-Faced Player in Damage and Repair. Ciryam P, Gerzanich V, Simard JM. J Neurotrauma. 2023 Nov;40(21-22):2249-2269. doi: 10.1089/neu.2023.0135. Epub 2023 Aug 10. PMID: 37166354; PMCID: PMC10649197.
2. Interleukin-6 as a prognostic biomarker of clinical outcomes after traumatic brain injury: a systematic review. Ooi SZY, Spencer RJ, Hodgson M, Mehta S, Phillips NL, Preest G, Manivannan S, Wise MP, Galea J, Zaben M. Neurosurg Rev. 2022 Oct;45(5):3035-3054. doi: 10.1007/s10143-022-01827-y. Epub 2022 Jul 6. PMID: 35790656; PMCID: PMC9256073.
3. Traumatic Brain Injury and Risk of Neurodegenerative Disorder. Brett BL, Gardner RC, Godbout J, Dams-O’Connor K, Keene CD. Biol Psychiatry. 2022 Mar 1;91(5):498-507. doi: 10.1016/j.biopsych.2021.05.025. Epub 2021 Jun 2. PMID: 34364650; PMCID: PMC8636548.
4. Role of IL-6 in the regulation of neuronal development, survival and function. Kummer KK, Zeidler M, Kalpachidou T, Kress M. Cytokine. 2021 Aug;144:155582. doi: 10.1016/j.cyto.2021.155582. Epub 2021 May 29. PMID: 34058569.

 

Testing Cytotoxic Activity of Drug Candidates with EZ4U – MTT Assay 

Globally, 19.3 million new cancer cases and nearly 10.0 million cancer deaths have been reported in 2020  (1). Despite the significant advances in cancer treatment, several challenges still remain e.g. avoiding  resistance or cytotoxic effects.  In search of more tolerable and effective drugs, a group of drug candidates have emerged in cancer research that target unique cell death mechanisms.

Ferroptosis is a form of regulated iron-dependent type of cell death that is driven by lipid peroxidation. It offers a new approach for developing novel drugs for cancer treatment. In the last years, several novel compounds capable of inducing ferroptosis in cancer have been investigated. Bernkop-Schnürch et al., recently evaluated the inhibitory effects of different methoxylated complexes on proliferation, migration, and metabolic activity in human breast cancer and leukemia cells. The researchers identified one promising lead compound for the design of a drug candidate that is capable of inducing ferroptosis in cancer cells (2). Read more: Design, Synthesis, Electrochemical, and Biological Evaluation of Fluorescent Chlorido[N,N‘-bis(methoxy/hydroxy)salicylidene-1,2-bis(4-methoxyphenyl)ethylenediamine]iron(III) Complexes as Anticancer Agents.

The Biomedica EZ4U Cell Proliferation and Cytotoxicity Assay (cat. no. BI-5000) was used in this study to evaluate the cytotoxic activity of the novel compounds. The assay is a modified MTT assay (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide) that detects the reduction of tetrazolium salts into yellow-colored formazan derivatives by functional mitochondria.

Brief, the metabolic activity was analysed by seeding cells in microtiter plates and culturing them at 37°C for 3 days in a 5% CO₂/95% air atmosphere. Thereafter, 20µl of the EZ4U substance was added to each well and the color change was measured on a microtiter plate reader at a wavelength of 450 nm with 620 nm as a reference.

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

• SAFE – non-radioactive & non-toxic 
• CONVENIENT –  single-step incubation for use on living cells
• TRUSTED – widely cited in over 260 publications

 

Download the BROCHURE – EZ4U cell proliferation and cytotoxicity assay and the PROTOCOL BOOKLET

 

 

Literature

1. Global Cancer Statistics 2020: GLOBOCAN Estimates of Incidence and Mortality Worldwide for 36 Cancers in 185 Countries. Sung H, Ferlay J, Siegel RL, Laversanne M, Soerjomataram I, Jemal A, Bray F.CA Cancer J Clin. 2021 May;71(3):209-249. doi: 10.3322/caac.21660. Epub 2021 Feb 4. PMID: 33538338.
2. Design, Synthesis, Electrochemical, and Biological Evaluation of Fluorescent Chlorido[N,N‘-bis(methoxy/hydroxy)salicylidene-1,2-bis(4-methoxyphenyl)ethylenediamine]iron(III) Complexes as Anticancer Agents. Bernkop-Schnürch AD, Chavooshi D, Descher HA, Leitner D, Talasz H, Hermann M, Wurst K, Hohloch S, Gust R, Kircher B. J Med Chem. 2023 Dec 14;66(23):15916-15925. doi: 10.1021/acs.jmedchem.3c01359. Epub 2023.

Machine Learning for Bone Biomarker Profiling in Rheumatoid Arthritis

Rheumatoid arthritis (RA) is a chronic, progressive inflammatory disorder which can lead to severe joint damage and disability. In 2019, an estimated 18 million people worldwide were living with this disease (1). Untreated RA can lead to destruction of the joints as well as heart, lung or nervous system problems (2). Skeletal bone loss, referred to as osteopenia or osteoporosis, is a key feature of RA. 

Sclerostin and Dickkopf-1 (DKK-1) are Wnt signaling proteins that are secreted by osteocytes, bone cells embedded in the bone matrix. They are inhibitors of bone formation and play a key role in the pathogeneses of systemic and localized bone loss in RA (3, 4). Serum levels of Sclerostin and DKK-1 have shown to be elevated in patients with RA compared to controls and correlate with bone erosions and inflammation (4, 5, 6). Findings in mice have demonstrated that DKK-1 triggers inflammatory bone degradation and neutralization of DKK-1 protects from systemic bone loss during inflammation (7). Interestingly, blocking the bone destruction molecule Sclerostin with an anti-sclerostin antibody has shown to be effective for the treatment of osteoporosis but may not be safe for patients suffering from inflammatory RA: in a rodent RA model, Weymeyer et al. demonstrated that Sclerostin inhibition did not stop bone loss and worsened clinical RA outcome by promoting TNF-dependent inflammatory joint destruction (8).

Controlling inflammation by biological therapies targeting pro-inflammatory cytokines has shown to have a positive effect in RA patients (5). Interleukin-6 is a key immunomodulatory cytokine that plays an important role in the development of RA. Inhibition of IL-6 has proven to be effective in treating patients with RA (9). A study by Briot et al showed that DKK-1 levels decreased in RA patients treated with an anti-IL-6 inhibitor (6).

Machine Learning for Bone Biomarker Profiling in Rheumatoid Arthritis

A recent cross-sectional study by Adami G et al., with over 1800 enrolled participants diagnosed with RA, Psoriatic Arthritis (PsA), and Systemic Sclerosis (SSc), employed machine learning techniques to assess the capability of biomarker profiles in differentiating RA patients from individuals with PsA and SSc. The Wnt signaling antagonists Sclerostin and Dickkopf-1 (DKK-1) were among the biomarkers measured. The study provided an in-depth understanding into the bone signature of RA that is marked by changes in bone mineral density and by unique biomarker profiles (6). Serum Sclerostin and DKK-1 levels were measured with ELISA assay kits from Biomedica.

SCLEROSTIN ELISA (#BI-20492) and DKK-1 (#BI-20403) ELISA kits

Biomedica´s Sclerostin ELISA Assay

• TRUSTED – cited in more than 290 publications!
• QUALITY – validated according to international guidelines
• EFFICIENT – only 20µl sample / well
• CONVENIENT – ready to use standards and controls included

 

Biomedica´s DKK-1 ELISA assay 

• TRUSTED – cited in more than 180 publications!
• QUALITY – validated according to international guidelines
• EFFICIENT – only 20µl sample / well
• CONVENIENT – direct measurement – no sample pre-dilution. Ready to use standards and controls included

 

Also available from Biomedica : Bioactive Sclerostin ELISA (cat. no. BI-20472), Interleukin-6 ELISA (BI-IL6)

 

Complete ready-to use ELISA kits

 

Literature

1. GBD 2019: Global burden of 369 diseases and injuries in 204 countries and territories, 1990–2019: a systematic analysis for the Global Burden of Disease Study 2019. https://vizhub.healthdata.org/gbd-results
2. WHO- Rheumatoid arthritis, June 2023
3. Wnt Signaling and Biological Therapy in Rheumatoid Arthritis and Spondyloarthritis. Cici D, Corrado A, Rotondo C, Cantatore FP. Int J Mol Sci. 2019 Nov 7;20(22):5552. doi: 10.3390/ijms20225552. PMID: 31703281; PMCID: PMC6888549.
4. Study of correlation of level of expression of Wnt signaling pathway inhibitors sclerostin and dickkopf-1 with disease activity and severity in rheumatoid arthritis patients. Singh A, Gupta MK, Mishra SP.Drug Discov Ther. 2019;13(1):22-27. doi: 10.5582/ddt.2019.01011. PMID: 30880318.
5. The effect of tocilizumab on bone mineral density, serum levels of Dickkopf-1 and bone remodeling markers in patients with rheumatoid arthritis. Briot K, Rouanet S, Schaeverbeke T, Etchepare F, Gaudin P, Perdriger A, Vray M, Steinberg G, Roux C. Joint Bone Spine. 2015 Mar;82(2):109-15. doi: 10.1016/j.jbspin.2014.10.015. Epub 2014 Dec 31. PMID: 25557658.
6. Machine learning to characterize bone biomarkers profile in rheumatoid arthritis. Adami G, Fassio A, Rossini M, Benini C, Bixio R, Rotta D, Viapiana O, Gatti D. Front Immunol. 2023 Nov 9;14:1291727. doi: 10.3389/fimmu.2023.1291727. PMID: 38022514; PMCID: PMC10665911.
7. Neutralisation of Dkk-1 protects from systemic bone loss during inflammation and reduces sclerostin expression. Heiland GR, Zwerina K, Baum W, Kireva T, Distler JH, Grisanti M, Asuncion F, Li X, Ominsky M, Richards W, Schett G, Zwerina J. Ann Rheum Dis. 2010 Dec;69(12):2152-9. doi: 10.1136/ard.2010.132852. Epub 2010 Sep 21. PMID: 20858621.
8. Sclerostin inhibition promotes TNF-dependent inflammatory joint destruction. Wehmeyer C, Frank S, Beckmann D, Böttcher M, Cromme C, König U, Fennen M, Held A, Paruzel P, Hartmann C, Stratis A, Korb-Pap A, Kamradt T, Kramer I, van den Berg W, Kneissel M, Pap T, Dankbar B. Sci Transl Med. 2016 Mar 16;8(330):330ra35. doi: 10.1126/scitranslmed.aac4351. Epub 2016 Mar 16. PMID: 27089204.
9. Targeting IL-6 or IL-6 Receptor in Rheumatoid Arthritis: What Have We Learned? Avci AB, Feist E, Burmester GR. BioDrugs. 2024 Jan;38(1):61-71. doi: 10.1007/s40259-023-00634-1. Epub 2023 Nov 21. PMID: 37989892; PMCID: PMC10789669.

 

The Biomedica SCLEROSTIN ELISA Assay Kit (# BI-20492) was utilized in a recent publication assessing the associations between serum and bone sclerostin levels and biomarkers of bone turnover and bone histomorphometry. Read more: Sclerostin, Osteocytes, and Wnt Signaling in Pediatric Renal Osteodystrophy.

Sclerostin (SOST) ELISA  (cat. no. BI-20492)

• Most referenced Sclerostin ELISA in over 290 citations
• Low sample volume – 20µl / well
• Validation following international guidelines

 

Sclerostin a biomarker in renal pediatric bone disease

Sclerostin, Osteocytes, and Wnt Signaling in Pediatric Renal Osteodystrophy. Laster M. et al., Nutrients. 2023 Sep 25;15(19):4127. doi: 10.3390/nu15194127. PMID: 37836411; PMCID: PMC10574198 . link to full text

Abstract

The pathophysiology of chronic kidney disease-mineral and bone disorder (CKD-MBD) is not well understood. Specific factors secreted by osteocytes are elevated in the serum of adults and pediatric patients with CKD-MBD, including FGF-23 and sclerostin, a known inhibitor of the Wnt signaling pathway. The molecular mechanisms that promote bone disease during the progression of CKD are incompletely understood. In this study, we performed a cross-sectional analysis of 87 pediatric patients with pre-dialysis CKD and post-dialysis (CKD 5D). We assessed the associations between serum and bone sclerostin levels and biomarkers of bone turnover and bone histomorphometry. We report that serum sclerostin levels were elevated in both early and late CKD. Higher circulating and bone sclerostin levels were associated with histomorphometric parameters of bone turnover and mineralization. Immunofluorescence analyses of bone biopsies evaluated osteocyte staining of antibodies towards the canonical Wnt target, β-catenin, in the phosphorylated (inhibited) or unphosphorylated (active) forms. Bone sclerostin was found to be colocalized with phosphorylated β-catenin, which suggests that Wnt signaling was inhibited. In patients with low serum sclerostin levels, increased unphosphorylated “active” β-catenin staining was observed in osteocytes. These data provide new mechanistic insight into the pathogenesis of CKD-MBD and suggest that sclerostin may offer a potential biomarker or therapeutic target in pediatric renal osteodystrophy.

 

Related Literature

FGF-23 and sclerostin in serum and bone of CKD patients. Lima F, Monier-Faugere MC, Mawad H, David V, Malluche HH. Clin Nephrol. 2023 May;99(5):209-218. doi: 10.5414/CN111111. PMID: 36970967; PMCID: PMC10286735. (Biomedica Sclerostin ELISA Assay Kit, cat. no. BI-20492 citation)

Sclerostin and Dickkopf-1 in renal osteodystrophy. Cejka D, Herberth J, Branscum AJ, Fardo DW, Monier-Faugere MC, Diarra D, Haas M, Malluche HH. Clin J Am Soc Nephrol. 2011 Apr;6(4):877-82. doi: 10.2215/CJN.06550810. Epub 2010 Dec 16. PMID: 21164019; PMCID: PMC3069382. (Biomedica Sclerostin ELISA Assay Kit, cat. no. BI-20492 citation)

Bone Disorders in Pediatric Chronic Kidney Disease: A Literature Review. Capossela L, Ferretti S, D’Alonzo S, Di Sarno L, Pansini V, Curatola A, Chiaretti A, Gatto A.Biology (Basel). 2023 Nov 2;12(11):1395. doi: 10.3390/biology12111395. PMID: 37997994; PMCID: PMC10669025.

 

Biomarkers have a longstanding role as indicators of biological changes. In drug development they serve dual purposes: predicting drug efficacy and identifying potential drug toxicity. Cardiotoxicity in one of the causes why preclinical safety tests fail during drug development. Therefore, monitoring cardiac toxicity with biomarkers is an essential part during drug development.

Cardiac Safety Biomarkers for Preclinical Cardiotoxicity Testing

The cardiac biomarkers NT-proANP and NT-proBNP

The biomarkers NT-proANP and NT-proBNP are cardiac hormones released in response to the stretching of the heart muscle . They belong to the group of natriuretic peptides and serve as cardiac biomarkers in both human and preclinical settings (1-4).

Biomedica offers reliable ELISA kits for measuring NT-proBNP and NT-proANP in both human and rodent samples.

Example of a Biomedica ELISA kit

 

Rat NT-proBNP ELISA – NEW

• Product code: BI-1204R
• Sample type: rat serum, plasma
• Sample volume: 10 µL/well
• Sensitivity: LOD 21 pg/ml
• Standard curve range: 0 – 3,200 pg/ml
• Specificity: Endogenous (natural) and recombinant human NT-proBNP (1-76)
• Assay time: 3.5 hours
• Reference value information for rat samples is provided. Instructions for use
• Citation click here (5)

 

also available: NT-proBNP ELISA (cat.no. SK-1204) – for human serum and plasma samples

• CE marked for IVD use in the EU
• Proficiency testing , saliva protocol
• Widely cited

 

NT-proANP ELISA

• Product code: BI-20892
• Sample types: Serum, plasma, urine, cell culture supernatant (human, rat, mouse, rabbit samples)

Due to the high sequence homology of NT-proANP among species the assay has been used not only in human samples but also in rodent (rat, mouse) rabbit, dog samples

• Sample volume: 10 µL/well
• Sensitivity: LOD 0.05 nmol/l (= 0.64 ng/ml)
• Standard curve range: 0 – 10 nmol/l (= 0 – 127 ng/ml)
• Specificity: endogenous (natural) and recombinant human NT-proANP (equivalent to proANP 1-98)
• There is high cross-reactivity between animal species.
• Assay time: 3.5 hours
• Citations all , citations with use of rat/mouse samples
• Instructions for use 

 

 

NT-proANP – a marker of drug-induced hypertrophy in rats

The validity of NT-proANP measurements as a tool for the detection of cardiovascular disorders in rats was demonstrated in an interlaboratory comparative study using Biomedica’s NT-proANP ELISA assay (Cat. No. BI-20892). Rat samples were analyzed and evaluated among 5 different laboratories by the PSTC-CHWG (Predictive Safety Testing Consortium and Cardiac Hypertrophy Working Group). The results demonstrated that the Biomedica NT-proANP ELISA assay (#BI-20892) is robust and technically adequate for the detection of serum NT-proANP levels in SD rats (6, 7).

 

Literature

1. Atrial and brain natriuretic peptides: Hormones secreted from the heart. Nakagawa Y, Nishikimi T, Kuwahara K. Peptides. 2019 Jan;111:18-25. doi: 10.1016/j.peptides.2018.05.012. Epub 2018 May 31. PMID: 29859763.
2. Cardiac natriuretic peptides. Goetze JP, Bruneau BG, Ramos HR, Ogawa T, de Bold MK, de Bold AJ. Nat Rev Cardiol. 2020 Nov;17(11):698-717. doi: 10.1038/s41569-020-0381-0. Epub 2020 May 22. PMID: 32444692.
3. Serum Natriuretic Peptides as Differential Biomarkers Allowing for the Distinction between Physiologic and Pathologic Left Ventricular Hypertrophy . Dunn, M.E., Manfredi, T.G., Agostinucci, K.,  Engle, S.K., Powe J., King, N.M.P., Rodriguez, L.A, Gropp, K.E., Gallacher, M., Vetter, F.J.,  More, V., Shimpi, P.,  Serra, D., Colton, H.M., for The Cardiac Hypertrophy Working Group of the Predictive Safety Testing Consortium . 2017. Toxicol Pathol 45(2): 334-352. PMID: 27102652
4. Natriuretic Peptides as Cardiovascular Safety Biomarkers in Rats: Comparison With Blood Pressure, Heart Rate, and Heart Weight . Engle, S.K., and Watson, D.E.  2016. Toxicol Sci 149: 458 – 472. PMID: 26609138.
5. Leucine Supplementation Improves Diastolic Function in HFpEF by HDAC4 Inhibition. Alves PKN, Schauer A, Augstein A, Männel A, Barthel P, Joachim D, Friedrich J, Prieto ME, Moriscot AS, Linke A, Adams V Cells. 2023 Nov 2;12(21):2561. doi: 10.3390/cells12212561. PMID: 37947639; PMCID: PMC10648219.
6. 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.J Pharmacol Toxicol Methods. 2016; 77:58-65. doi: 10.1016/j.vascn.2015.10.002. PMID: 26516096.
7. 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 et al., Toxicol Pathol. 2017; 45(2):344-352.

 

 

 

 

 

Kidney disease is estimated to affect more than 800 million people worldwide (1). Taking action for prevention, early diagnosis and treatment can reduce the risk and the burden of kidney disease.

Role of the kidneys, kidney disease and risk factors

Kidneys are vital organs that regulate fluid balance, blood pressure and produce hormones that stimulate the production of red blood cells . Kidney disease is a condition in which kidneys lose their ability to effectively filter waste products and excess fluids from the blood. Kidney disease commonly leads to a decline in kidney function that may lead to kidney failure, characterized by the complete loss of kidney function. At this stage dialysis or kidney transplantation become the only treatment option.

Kidney problems can emerge suddenly (acute) or gradually over time (chronic). Various conditions, diseases and medications can contribute to acute and chronic kidney problems.  Chronic kidney disease (CKD) is characterized by a prolonged period of kidney abnormalities that last for more than three months (2), whereas acute kidney disease – acute kidney injury (AKI) is characterized by a sudden loss of excretory kidney function (3).

Other forms of kidney disease include polycystic kidney disease (PKD) a genetic disorder that leads to kidney enlargement and impaired kidney function over time and glomerulonephritis (GN). GN is a group of diseases characterized by inflammation of the glomeruli, the filtration units of the kidney (4).

The most common risk factors for the development and progression of CKD are diabetes and high blood pressure. Managing blood sugar and blood pressure can help keep kidneys healthy. Other risk factors of CKD include heart disease, obesity, family history and genetic background as well as age, smoking and nutrition (5).

Consequences of kidney disease include heart disease, high blood pressure, bone and mineral disorders, and anemia (6).

Keep Your Kidneys Healthy

Regular exercise, weight control, a balanced diet (7) and sufficient fluid intake are only some of the ways to keep your kidney healthy.

World Kidney Day – about kidney health download here and check out the 6-Step Guide to Protecting Kidney Health here .

BIOMEDICA – Biomarker ELISA kits for clinical research in kidney disease

• HIGH QUALITY ASSAYS – Fully validated according to international quality guidelines
• TRUSTED – Widely cited in over 1500 publications

 

ELISA kits for: FGF23 (Fibroblast growth factor 23), Vanin-1, Endostatin, Sclerostin, Osteoprotegerin (OPG), Angiopoietin-2 (ANG-2), IL-6 (Interleukin-6) , VEGF (Vascular Endothelial Growth Factor)

Check out our brochure on Biomarkers in Clinical Nephrology  

 

 

Literature

1. Epidemiology of chronic kidney disease: an update 2022. Kidney Int Suppl (2011). Kovesdy CP. 2022 Apr;12(1):7-11. doi: 10.1016/j.kisu.2021.11.003. Epub 2022 Mar 18. PMID: 35529086; PMCID: PMC9073222.
2. Chronic Kidney Disease Diagnosis and Management: A Review. Chen TK, Knicely DH, Grams ME. JAMA. 2019 Oct 1;322(13):1294-1304. doi: 10.1001/jama.2019.14745. PMID: 31573641; PMCID: PMC7015670.
3. Acute kidney injury. Kellum JA, Romagnani P, Ashuntantang G, Ronco C, Zarbock A, Anders HJ. Nat Rev Dis Primers. 2021 Jul 15;7(1):52. doi: 10.1038/s41572-021-00284-z. PMID: 34267223.
4. Glomerulonephritis: immunopathogenesis and immunotherapy. Anders HJ, Kitching AR, Leung N, Romagnani P.Nat Rev Immunol. 2023 Jul;23(7):453-471. doi: 10.1038/s41577-022-00816-y. Epub 2023 Jan 12. PMID: 36635359; PMCID: PMC9838307.
5. Risk factors for chronic kidney disease: an update. Kazancioğlu R Kidney Int Suppl (2011). 2013 Dec;3(4):368-371. doi: 10.1038/kisup.2013.79. PMID: 25019021; PMCID: PMC4089662.
6. Cardiorenal Syndrome and the Role of the Bone-Mineral Axis and Anemia. Charytan DM, Fishbane S, Malyszko J, McCullough PA, Goldsmith D Am J Kidney Dis. 2015 Aug;66(2):196-205. doi: 10.1053/j.ajkd.2014.12.016. Epub 2015 Feb 26. PMID: 25727384; PMCID: PMC4516683.
7. The Effect of Diet on the Survival of Patients with Chronic Kidney Disease. Rysz J, Franczyk B, Ciałkowska-Rysz A, Gluba-Brzózka A.Nutrients. 2017 May 13;9(5):495. doi: 10.3390/nu9050495. PMID: 28505087; PMCID: PMC5452225.

 

 

 

Chronic kidney disease is a progressive condition that affects >10% of the general population worldwide (1). Current clinical biomarkers prove effective at advanced stages of renal impairment, limiting the timely initiation of potentially successful therapeutic interventions. There is an unmet need for more refined biomarkers capable of detecting CKD at earlier stages, thereby enhancing the prospects for patients’ outcomes.

In the last decade, the advancement in the fields of genomics, proteomics, and metabolomics have led to the identification of potential biomarker candidates that may offer important diagnostic and prognostic information in patients suffering from kidney diseases (2).

Among the current established biomarkers such as serum creatinine, albuminuria, and proteinuria, novel biomarkers for kidney diseases could potentially provide additional prognostic information. They could help to predict treatment response in various clinical settings such as acute kidney injury, transplant rejection or glomerulopathies (2).

Emerging Biomarkers in Kidney Disease

Biomedica offers a range of ELISA assay kits to reliably detect biomarkers in blood samples of patients with kidney diseases.

Endostatin, Vanin-1, Periostin, FGF23, IL-6 and more…

 

 

ENDOSTATIN – a potential biomarker of renal fibrosis, chronic kidney disease (CKD), prognostic marker in acute kidney injury (AKI)

Endostatin is an extracellular matrix protein which is expressed in patients during the progression of renal fibrosis. The significant increase of serum Endostatin levels may be due to the enhanced degradation of the extracellular matrix in patients with chronic kidney disease (3, 4). Endostatin has also been studied as a prognostic marker in patients with acute kidney injury (AKI) (5) and is independently associated with incident cardiovascular events in CKD patients (6).

Endostatin can reliably be quantified in serum, plasma and urine samples:

• Endostatin ELISA | BI-20742
• Endostatin ELISA (Mouse/Rat) | BI-20742MR  (7)

 

VANIN-1 – a potential biomarker of acute kidney injury and drug induced renal injury

Vascular non-inflammatory molecule-1 (Vanin-1) is highly expressed in the kidney (8) and has been proposed as a marker in acute kidney injury and drug induced renal injury (9).  Vanin-1 has been identified as a marker of kidney damage as shown n a rat model of type 1 diabetic nephropathy (10).

Urinary Vanin-1  has been investigated in children with renal fibrosis (11) and as a predictor of acute pyelonephritis in young children with urinary tract infection (12). A recent study investigated the role of urinary Vanin-1 in kidney transplant recipients (13).

Vanin-1 can easily be measured with a conventional ELISA assay:

• Vanin-1 (urine) ELISA | BI-VAN1U
• Vanin-1 ELISA (Mouse/Rat) ELISA | BI-VAN1MR        

 

PERIOSTIN – a potential early biomarker of renal tubular injury

Periostin is a matricellular protein that is involved in tissue remodeling and wound healing. Studies have demonstrated that the expression of Periostin in the kidney correlates with the degree of interstitial fibrosis and a decline in kidney function (15). Elevated urine Periostin levels were found in patients with type 2 diabetes which were present before the onset of microaluminuria. The authors proposed that urinary Periostin could be an early biomarker of renal tubular injury (16).

Periostin can reliably be measured in serum, plasma, and urine samples with a fully validated ELISA assay (17).

• Periostin ELISA | BI-20422
• Periostin ELISA (Mouse/Rat) | BI-20433MR

 

FGF23 – a potential early biomarker cardiovascular events in patients with renal-cardiovascular disease

Fibroblast growth factor 23 (FGF23) is an endocrine hormone that regulates phosphate homeostasis by modulating renal phosphate reabsorption in the kidney. Circulating FGF23 increases with declining kidney function and high FGF23 and phosphate levels are related to cardiovascular disease and mortality (18, 19). 

• FGF23  (C-terminal) multimatrix ELISA | BI-20702
• FGF23 Intact ELISA | BI-20700

 

IL-6 – a biomarker of inflammation

 Interleukin-6 (IL-6) is a cytokine that plays a crucial role in inflammation and in the regulation of immune response. It is a signaling molecule that is involved in various physiological processes, including the activation of immune cells and the coordination of responses to infections or injury. IL-6 is implicated in diseases where inflammation is a prominent feature (20).

•  IL-6 ELISA | BI-IL6
• HIGHLY SENSITIVE – measurable values in serum and plasma samples
• EASY – ready to use calibrators and controls

 

 

Literature

1.  Epidemiology of chronic kidney disease: an update 2022. Kovesdy CP. Kidney Int Suppl (2011). 2022. 12(1):7-11. doi: 10.1016/j.kisu.2021.11.003. PMID: 35529086; PMCID: PMC9073222.
2. Emerging Biomarkers for Early Detection of Chronic Kidney Disease. Mizdrak M, Kumrić M, Kurir TT, Božić J. J Pers Med. 2022. 12(4):548. doi: 10.3390/jpm12040548. PMID: 35455664.
3. A defective angiogenesis in chronic kidney disease. Futrakul N, Butthep P, Laohareungpanya N, Chaisuriya P, Ratanabanangkoon K. Ren Fail. 2008. 30(2):215-7. doi: 0.1080/08860220701813335. PMID: 18300124.
4. Endostatin in Renal and Cardiovascular Diseases. Li M, Popovic Z, Chu C, Krämer BK, Hocher B., Kidney Dis (Basel). 2021.9;7(6):468-481. doi: 10.1159/000518221. PMID: 34901193.
5. Prognostic value of dynamic plasma endostatin for the prediction of mortality in acute kidney injury: A prospective cohort study. Jia HM, Zheng Y, Han Y, Ma WL, Jiang YJ, Zheng X, Guo SY, Zhang TE, Li WX., J Int Med Res. 2020.  48(7):300060520940856. PMID: 32691651.
6. Endostatin in chronic kidney disease: Associations with inflammation, vascular abnormalities, cardiovascular events and survival. Kanbay M, Afsar B, Siriopol D, Unal HU, Karaman M, Saglam M, Gezer M, Taş A, Eyileten T, Guler AK, Aydin İ, Oguz Y, Tarim K, Covic A, Yilmaz MI. Eur J Intern Med. 2016. 33:81-7. doi: 10.1016/j.ejim.2016.06.033. PMID: 27394925.
7. Validation of an enzyme-linked immunosorbent assay (ELISA) for quantification of endostatin levels in mice as a biomarker of developing glomerulonephritis. Wallwitz J, Aigner P, Gadermaier E, Bauer E, Casanova E, Bauer A, Stoiber D. PLoS One. 2019. 14(8):e0220935. doi: 10.1371/journal.pone.0220935. PMID: 31404120.
8. Chemical biology tools to study pantetheinases of the vanin family. Schalkwijk, J, Jansen, P. Biochem Soc Trans. 2014. 42, 1052–1055.
9. Urinary Vanin-1 As a Novel Biomarker for Early Detection of Drug-Induced Acute Kidney Injury. Hosohata K et al.J  Pharm  Exp Ther. 2012. 341, 656–662.
10. Proteomic identification of vanin-1 as a marker of kidney damage in a rat model of type 1 diabetic nephropathy. Fugmann T et al., Kidney Int. 2011. 80, 272–281.
11. The Usefulness of Vanin-1 and Periostin as Markers of an Active Autoimmune Process or Renal Fibrosis in Children with IgA Nephropathy and IgA Vasculitis with Nephritis-A Pilot Study. Mizerska-Wasiak M, Płatos E, Cichoń-Kawa K, Demkow U, Pańczyk-Tomaszewska M. J Clin Med. 2022. 11(5):1265. doi: 10.3390/jcm11051265. PMID: 35268356. 
12. Urinary vanin-1 for predicting acute pyelonephritis in young children with urinary tract infection: a pilot study. Krzemień G, Pańczyk-Tomaszewska M, Górska E, Szmigielska A. Biomarkers. 2021. 26(4):318-324.
13. Urinary vanin-1, tubular injury, and graft failure in kidney transplant recipients. Alkaff FF, Kremer D, Niekolaas TM, van den Born J, Rimbach G, Tseng TL, Berger SP, Bakker SJL, de Borst MH. Sci Rep. 2024. 4(1):2283. doi: 10.1038/s41598-024-52635-x. PMID: 38280883.
14. Development of an immunoassay that reveals altered uninary Vanin-1 in human with kidney disease. Wallwitz, J., Eichinger, B., Berg, G., Gadermaier, E., Himmler, G. 2018. Nephrology Dialysis Transplantation, Volume 33, Issue suppl_1, Page i126. 
15. Periostin in the Kidney. Wallace DP et al., Adv Exp Med Biol. 2019, 1132:99-112.
16. Periostin as a tissue and urinary biomarker of renal injury in type 2 diabetes mellitus. Satirapoj B et al., PLoS One. 2015, 17;10(4):e0124055.
17. Characterization of a sandwich ELISA for the quantification of all human periostin isoforms. Gadermaier E et al., J Clin Lab Anal. 2018, 32(2):e22252.
18. Higher fibroblast growth factor-23 increases the risk of all-cause and cardiovascular mortality in the community. Ärnlöv J, Carlsson AC, Sundström J, Ingelsson E, Larsson A, Lind L, Larsson TE.  Kidney Int. 2013. 83(1):160-6. doi: 10.1038/ki.2012.327. PMID: 22951890. 
19. Fibroblast growth factor-23, cardiovascular prognosis, and benefit of angiotensin-converting enzyme inhibition in stable ischemic heart disease. Udell JA, Morrow DA, Jarolim P, Sloan S, Hoffman EB, O’Donnell TF, Vora AN, Omland T, Solomon SD, Pfeffer MA, Braunwald E, Sabatine MS. J Am Coll Cardiol. 2014. 10;63(22):2421-8, doi: 10.1016/j.jacc.2014.03.026. PMID: 24727254; PMCID: PMC4213068.
20. Interleukin 6 and Cardiovascular Outcomes in Patients With Chronic Kidney Disease and Chronic Coronary Syndrome. Batra G, Ghukasyan Lakic T, Lindbäck J, Held C, White HD, Stewart RAH, Koenig W, Cannon CP, Budaj A, Hagström E, Siegbahn A, Wallentin L; STABILITY Investigators. JAMA Cardiol. 2021. 6(12):1440-1445. doi: 10.1001/jamacardio.2021.3079. PMID: 34431970; PMCID: PMC8387946.
21.  

 

 

 

 

 

 

 

 

 

 

 

 

Rare Disease Day is a global initiative held on the last day of February. It raises awareness for rare diseases to improve accessibility to medical treatment and representation for individuals diagnosed with rare diseases. It is estimated that around 300 million people worldwide are living with rare diseases.

Rare metabolic bone diseases are caused by genetic disorders that may directly or indirectly have an impact on bone structure or function (1). Other factors like hormones, tumors, diet or certain medications may also lead to abnormal growth and development of the skeleton. Some of the diseases are inherited many caused by genetic mutations. Other bone disorders are not inherited and can develop after birth.In some cases, the precise cause remains unknown.

Rare bone diseases

Rare bone diseases account for 5% of all birth defects and are an important cause of disability worldwide, yet they remain a difficult group of conditions to treat (2). It is estimated that more than 400 developmental abnormalties of the skeletal system exist (3).

The main rare metabolic bone diseases include Hypophosphatemia, Osteogenesis Imperfecta, Tumor-Induced Osteomalacia, X-Linked Hypophashatemia, and other Rare Bone Diseases (Fibrous Dysplasia, Osteopetrosis, High Bone Mass..) (4).

Biomarkers in Rare Bone Diseases

Biomarkers in Rare Bone Diseases provide a way to accelerate medical research by providing valuable insights into disease mechanisms. They play an important role in monitoring disease progression, optimizing treatments. and developing novel therapies.

NT-proCNP  

C-natriuretic peptide (CNP) and its receptor, natriuretic peptide receptor-B (NPR-B) are important regulators of endochondral ossification and longitudinal bone growth (5, 6)  The discovery and understanding of their physiological functions in promoting longitudinal bone growth have created opportunities for a specific targeted strategy in achondroplasia, the most common form of human dwarfism (7).

RANKL and OPG

Receptor activator of nuclear factor (NF-kappaβ) ligand (RANKL), its cellular receptor, receptor activator of NF-kappaβ (RANK), and the decoy receptor osteoprotegerin (OPG) are part of a cytokine system that regulate bone formation and resorption (8) . Denosumab is a bone anti-resorptive drug, a monoclonal antibody that binds RANKL and disrupts bone resorption. It has been approved for the treatment of osteoporosis and other bone-related disorders. The use of Denosumab in pediatric patients with Osteogenesis Imperfecta (OI), a genetic bone disorder, also known as brittle bone disease, shows decreased fractures and improved bone growth (9). A clinical trial at the National Institutes of Health found that Denosumab, significantly reduced abnormal bone turnover in adults with fibrous dysplasia, a rare disease characterized by weak, oddly shaped, or broke bones.

SCLEROSTIN

Sclerostin is a secreted protein that decreases bone formation. It binds to LRP-5 receptor on the surface of osteoblasts and consequently interferes with WNT signalling. Genetic sclerostin deficiency leads to increased bone formation and sclerotic bone disorders. Sclerostin inhibition is being evaluated as a potential approach to increase bone mass in Osteogenesis Imperfecta (10).

FGF23

Fibroblast growth factor 23 (FGF23) is a hormone that is produced by bone. It regulates serum phosphate levels by suppressing phosphate reabsorption in the kidney. Excessive actions of FGF23 are responsible for different kinds of hypophosphatemic rickets as found in X-linked hypophosphatemia (XLH) and osteomalacia. XLH is characterized by deformities of the lower limb and short stature. An anti-fibroblast growth factor-23 (FGF23) monoclonal antibody (Burosumab) has been approved as a novel treatment for hypophopshatemic rickets (11).

Measurement of FGF23 is a critical tool to assist in the evaluation and diagnosis of hypophosphatemic conditions (12, 13).

The second most common genetic form of hypophosphatemic rickets after XLH, is autosomal-dominant hypophosphatemic rickets (ADHR). ADHR is caused by specific mutations in the FGF23 gene.

FGF23 can reliably be measured with an immunoassay (14).

FGF23 ELISAs (c-term FGF23, cat. no BI-20702) (intact FGF23, cat. no. BI-20700)

• MULTI-USE for serum and plasma samples
• TRUSTED in over 60 citations

 

FGF23 – Test

 

Biomarker ELISA assay kits from BIOMEDICA

Complete ready-to use ELISA kits 

 

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

• CONVENIENT – low sample volume – 20 µl / well
• RELIABLE – validated following international quality guidelines
• TRUSTED – widely cited in over 40 publications (click here for full list)
• MULTI-USE – for human serum and plasma samples; protocols for non-human samples (e.g. rat). 

 

Citations Selection of NT-proCNP ELISA citations related to skeletal disorders:

• Plasma C-Type Natriuretic Peptide: Emerging Applications in Disorders of Skeletal Growth. Espiner E et al.,Horm Res Paediatr. 2018. 90(6):345-357. doi: 10.1159/000496544. PMID: 30844819.
• Rats deficient C-type natriuretic peptide suffer from impaired skeletal growth without early death. Fujii T et al., PLoS One. 2018. 22;13(3):e0194812. doi: 10.1371/journal.pone.0194812. PMID: 29566041.
• Serum NT-proCNP levels increased after initiation of GH treatment in patients with achondroplasia/hypochondroplasia. Kubota T etal., Clin Endocrinol (Oxf). 2016 Jun;84(6):845-50. doi: 10.1111/cen.13025. Epub 2016 Feb 25. PMID: 26814021.
• Acromesomelic dysplasia, type maroteaux caused by novel loss-of-function mutations of the NPR2 gene: Three case reports. Wang W et al., 2016. Am J Med Genet A 170A, 426–434.
• Skeletal overgrowth syndrome caused by overexpression of C-type natriuretic peptide in a girl with balanced chromosomal translocation, t(1;2)(q41;q37.1). Ko J et al., 2015. Am J Med Genet A 167A, 1033–1038. 

 

RANKL ELISA (cat. no. BI-20462)

• Highly sensitive – measurable concentrations in healthy subjects
• Only assay that measures free, uncomplexed, soluble RANK Ligand
• TRUSTED in over 300 citations 

 

OPG ELISA (BI-20403)

• CONVENIENT- ready to use liquid calibrators and controls
• EFFICIENT – low sample volume 20µl / well
• TRUSTED in over 250 citations 

 

Literature 

1. Genetic approaches to metabolic bone diseases. Hannan FM, Newey PJ, Whyte MP, Thakker RV. Br J Clin Pharmacol. 2019. 85(6):1147-1160. doi: 10.1111/bcp.13803. PMID: 30357886; PMCID: PMC6533455.
2. The evolving therapeutic landscape of genetic skeletal disorders. Sabir, A.H., Cole, T. Orphanet J Rare Dis 14, 300 (2019).
3. Changes in skeletal dysplasia nosology. Jurcă MC, Jurcă SI, Mirodot F, Bercea B, Severin EM, Bembea M, Jurcă AD. Rom J Morphol Embryol. 2021. 2(3):689-696. doi: 10.47162/RJME.62.3.05. PMID: 35263396; PMCID: PMC9019670.
4. Atlas of rare genetic metabolic bone diseases. IOF, 2024.
5. Natriuretic peptide regulation of endochondral ossification. Evidence for possible roles of the C-type natriuretic peptide/guanylyl cyclase-B pathway. Yasoda A, Ogawa Y, Suda M, Tamura N, Mori K, Sakuma Y, Chusho H, Shiota K, Tanaka K, Nakao K J Biol Chem.1998. 8;273(19):11695-700. doi: 10.1074/jbc.273.19.11695. PMID: 9565590.
6. Cyclic GMP-dependent protein kinase II plays a critical role in C-type natriuretic peptide-mediated endochondral ossification. Miyazawa T, Ogawa Y, Chusho H, Yasoda A, Tamura N, Komatsu Y, Pfeifer A, Hofmann F, Nakao K. Endocrinology. 2002. 143(9):3604-10. doi: 10.1210/en.2002-220307. PMID: 12193576.
7. Optimal management of complications associated with achondroplasia. Ireland PJ, Pacey V, Zankl A, Edwards P, Johnston LM, Savarirayan R Appl Clin Genet. 2014. 7:117-25. doi: 10.2147/TACG.S51485. PMID: 25053890; PMCID: PMC4104450.
8. C-type natriuretic peptide regulates endochondral bone growth through p38 MAP kinase-dependent and -independent pathways. Agoston H, Khan S, James CG, Gillespie JR, Serra R, Stanton LA, Beier F BMC Dev Biol. 2007. 7:18. doi: 10.1186/1471-213X-7-18. PMID: 17374144; PMCID: PMC1847438.
9. Role of receptor activator of nuclear factor-kappaB ligand and osteoprotegerin in bone cell biology. Hofbauer LC, Heufelder AE. J Mol Med (Berl). 2001 Jun;79(5-6):243-53. doi: 10.1007/s001090100226. PMID: 11485016.
10. Effect of denosumab on the growing skeleton in osteogenesis imperfecta. Hoyer-Kuhn H, Semler O, Schoenau E.J Clin Endocrinol Metab. 2014. 99(11):3954-5. doi: 10.1210/jc.2014-3072. PMID: 25148238.
11. Effect of sclerostin inactivation in a mouse model of severe dominant osteogenesis imperfecta. Marulanda J, Tauer JT, Boraschi-Diaz I, Bardai G, Rauch F. Sci Rep. 2023. 13(1):5010. doi: 10.1038/s41598-023-32221-3. PMID: 36973504; PMCID: PMC10043013.
12. Hereditary Metabolic Bone Diseases: A Review of Pathogenesis, Diagnosis and Management. Charoenngam N, Nasr A, Shirvani A, Holick MF.Genes (Basel). 2022. 13(10):1880. doi: 10.3390/genes13101880. PMID: 36292765; PMCID: PMC9601711.
13. Determination of FGF23 Levels for the Diagnosis of FGF23-Mediated Hypophosphatemia. Hartley IR, Gafni RI, Roszko KL, Brown SM, de Castro LF, Saikali A, Ferreira CR, Gahl WA, Pacak K, Blau JE, Boyce AM, Salusky IB, Collins MT, Florenzano P. J Bone Miner Res. 2022. 37(11):2174-2185. doi: 10.1002/jbmr.4702. PMID: 36093861; PMCID: PMC9712269.
14. The Measurement and Interpretation of Fibroblast Growth Factor 23 (FGF23) Concentrations. Heijboer AC, Cavalier E Calcif Tissue Int. 2023. 112(2):258-270. doi: 10.1007/s00223-022-00987-9. PMID: 35665817; PMCID: PMC9859838.

 

 

 

Heart failure (HF) is a leading cause of death in individuals with chronic kidney disease (CKD). CKD is also known as a “silent killer” as many people live with the disease without developing symptoms. CKD is a progressive disease and it is estimated that around 10% of the adult general population worldwide is affected (1). A large number of individuals are undiagnosed as demonstrated in a study from the UK in 3200 participants aged over 60 years (2).  

FGF23 and Risk of Heart Failure

What is CKD?

Chronic kidney disease is a condition where kidney function gradually declines. The kidneys play a crucial role in filtering waste from the blood with the help of nephrons. A constant increase in damaged and non-functioning nephrons lead to the progression of the disease.

How does CKD cause Heart Failure?

Impaired kidney function places an additional strain on the heart, as it needs to pump harder to get blood to the kidneys. Changes in blood pressure is also a complication in CKD that can ultimately lead to heart disease. One suggested mechanism for the development of HF in CKD patients is linked to highly elevated levels of FGF23 (3).

What is the role of FGF23 in Heart Failure?

Fibroblast growth factor 23 (FGF 23) is a bone-derived hormone that regulates phosphate and vitamin D. In individuals with CKD, circulating FGF23 blood levels gradually increase with declining kidney function. In patients with end-stage renal disease, FGF23 levels are highly elevated up to 1000-fold above the normal range (4).

Numerous studies indicate that high FGF23 concentrations have a direct effect on the heart and the cardiovascular system. Elevated FGF23 levels have been linked to an increased risk of cardiovascular events, including heart failure, cardiovascular mortality, and left ventricular hypertrophy (5, 6). The precise mechanisms by which FGF23 impacts the heart are still being explored. One potential pathway could be linked to the discovery of FGF23 receptors expressed in the heart by cardiomyocytes. Specific binding of FGF23 to these receptors may directly induce hypertrophy. Additional pathways are under investigation.

In a recent large multicenter prospective cohort study in over 3500 participants, researchers investigated the relationship between elevated FGF23 levels and the risk of HF in individuals with varies HF subtypes. The authors concluded that elevated FGF23 levels were associated with increased risks for all HF subtypes highlighting the need for further research on FGF23 as a potential target in HF prevention in individuals with chronic kidney disease (3).

What are the methods for measuring FGF23?

Circulating blood FGF23 levels can be measured with immunoassays. A commonly employed technique is the ELISA assay (enzyme linked immunosorbent assay).

FGF23 ELISA assays employ specific antibodies designed to detect FGF23 present in the sample (serum / plasma). Currently, two commercially available assays are used for the measurement of circulating FGF23 in blood samples (7):

• ELISA for measuring Intact FGF23

The full length, biologically active form of the hormone is intact FGF23 (iFGF23), consisting of the complete and full-length FGF23 protein structure without enzymatical cleavage. The ELISA utilizes two antibodies that target the N-terminal part and the C-terminal domain of the FGF23 molecule, respectively.

 

 

• ELISA for measuring FGF23 C-terminal

C-terminal fragments of FGF23 (cFGF23) result from the enzymatic cleavage of the intact FGF23 molecule. The ELISA utilizes two antibodies that bind to epitopes that are located in the c-terminal domain of the FGF23 molecule.

 

 

OF NOTE: all FGF23 c-terminal assays that are commercially available detect both c-terminal FGF23 fragments as well as the intact FGF23 molecule (7).

 

BIOMEDICA offers two ELISA kits that reliably quantify FGF23 concentrations in human serum and plasma samples:

FGF23 – complete ready to use ELISA kits – User-Friendly and Reliable

 

• FGF23 intact ELISA (cat. no. BI-20700)
• FGF23 (C-terminal) ELISA (cat. no. BI-20702)

 

BIOMEDICA FGF23 intact ELISA assay (#BI-20700)

 

BIOMEDICA FGF23 multimatrix (C-terminal) ELISA assay (#BI-20702)

 

• Validated FGF23 ELISA kits according to international quality guidelines
• Numerous citations /references in top-ranking journals

 

FGF23 ELISA assays are Developed & Manufactured by Biomedica

Literature

1. Epidemiology of chronic kidney disease: an update 2022. Kovesdy CP. Kidney Int Suppl (2011). 2022. 12(1):7-11. PMID: 35529086; PMCID: PMC9073222
2. Prevalence of chronic kidney disease in the community using data from OxRen: a UK population-based cohort study. Hirst JAet al., Br J Gen Pract. 2020. 26;70(693):e285-e293. PMID: 32041766; PMCID: PMC7015167.
3. Chronic Renal Insufficiency Cohort (CRIC) study investigators. Fibroblast Growth Factor 23 and Risk of Heart Failure Subtype: The CRIC (Chronic Renal Insufficiency Cohort) Study. Leidner Aset al.,Kidney Med. 2023. 5(11):100723. PMID: 37915961; PMCID: PMC10616385.
4. Fibroblast growth factor-23 mitigates hyperphosphatemia but accentuates calcitriol deficiency in chronic kidney disease. Gutierrez O et al., Journal of the American Society of Nephrology. 2005;16(7):2205–2215. 
5. FGF23 and Cardiovascular Risk. Prié D. Ann Endocrinol (Paris). 2021. 82(3-4):141-143. PMID: 32950228.
6. Paracrine Effects of FGF23 on the Heart. Front Endocrinol (Lausanne). Leifheit-Nestler M, Haffner D 2018. 28;9:278. PMID: 29892269; PMCID: PMC5985311.
7. The Measurement and Interpretation of Fibroblast Growth Factor 23 (FGF23) Concentrations. Calcif Tissue Int. Heijboer AC, Cavalier E. 2023 Feb;112(2):258-270. PMID: 35665817; PMCID: PMC9859838.

 

 

The enzyme-linked immunosorbent assay (ELISA or EIA) is a laboratory method to detect and quantify the presence of a protein in biological samples (1, 2).

When selecting an ELISA kit, researchers are often confronted with the question which assay to choose of the many commercially available kits.

It can be a challenge! Here are a few hints that may help.

HOW TO CHOOSE THE RIGHT ELISA KIT

As a general rule, before purchasing an assay, always read the protocol booklet (instructions for use – IFU or package insert) in detail. This should ensure that the kit will be suitable for your requirements. Check out the following:

1. ANALYTE

Which protein biomarker will you be measuring? Be sure to use the correct term during your search. Some biomarkers have alternative names (e.g. Sclerostin or SOST ELISA (SOST is actually the name for the gene that encodes Sclerostin).

2. SPECIES – SPECIFICITY – CROSS REACTIVITY

Verify if the assay can be used in the respective model such as e.g. human, rat, or mouse. Due to high homology between species, some ELISA kits work both in humans and in different animal species. As an example the biomarker ELISA kit for NT-proANP was developed for human use but due to the high sequence homology between species, the kit is successfully used to measure NT-proANP as a cardiac safety biomarker in various animal models (rat, mouse, rabbit, monkey).

3. SAMPLE TYPE

What is the sample type (matrix) you´ll be using (e.g. serum, EDTA-plasma, heparin-plasma, citrate-plasma, cell culture supernatants, urine..) ?

Verify if the assay is compatible for your sample type: check the package insert and, if available, check if there are validation data showing results (often found on the manufacturers website).

Of note: analysis of some biomarkers in the “wrong” matrix can lead to “false” results due to a matrix effect.

4. SAMPLE VOLUME

Check the amount of sample required per well (calculate volume to measure your samples in duplicates). Low sample volumes and precious samples are often a selection criterium.  

5. SENSITIVTY – BIOMARKER CONCENTRATIONS TO BE EXPECTED

Before choosing an assay, look into the validation data of the kit (often documented in the ELISA protocol booklet or in the validation data files).  

Reference values and pathological values in serum and/or plasma of the biomarker of interest are sometimes documented as well. These data can be helpful in selecting an appropriate assay. In some cases samples may require a pre-dilution. Therefore, always verify if the dilution buffer (assay buffer) is included in the kit or ask the assay developer for their input.

Of note: assays offering high sensitivity offer a different dynamic range than assays with a lower sensitivity. The dynamic range of an assay indicates the range of concentrations over which an assay can accurately quantify the analyte.

6. ASSAY PERFORMANCE – ASSAY VALIDATION

Careful evaluation of the assay´s performance characteristics is important in selecting an ELISA kit.

Choose an assay that has gone through a rigorous validation process. Check out if you can find data on the following performance characteristics:

• Accuracy- detection of a protein biomarker in clinical samples.
• Dilution linearity and parallelism  – recovery of the analyte of interest in diluted samples
• Specificity & cross-reactivity – making sure that you detect only the analyte of interest
• Precision – within-run and in-between run precision – ensuring precise and reproducible results within an across assay lots
• Calibration – ensures consistent performance over the range of the assay of the calibration curve
• Sample stability – ensures the stability of the analyte of interest (e.g. exposure of real samples to multiple freeze-thaw cycles, stability at room temperature..).

 

7. KIT COMPONENTS

Verify if the contents of the ELISA kit includes all the necessary components e.g. controls, assay dilution buffer. Consider storage requirements such as temperature sensitivity and expiration date.

8. CITATIONS & REFERENCES

Check if there are citations on the manufacturers website for the specific ELISA kit. Look into publications and seek feedback from researchers who have used the assay you are considering.

9. PRODUCT ORIGIN

Verify if the kit supplier is the kit manufacturer. More and more kits are repacked and are sold under different names, although it is always the same kit.

ELISA kit manufacturers will more likely give you qualified support as they “know” their product (e.g. availability of additional calibrators, controls, buffers.., technical know-how on the kit..).

10. CUSTOMER SUPPORT

Verify if the kit provider can provide timely and helpful customer service.

 

Related Literature

1.Enzyme-Linked Immunosorbent Assay: Types and Applications. Hayrapetyan H, Tran T, Tellez-Corrales E, Madiraju C. Methods Mol Biol. 2023;2612:1-17. doi: 10.1007/978-1-0716-2903-1_1. PMID: 36795355.

2. Enzyme Immunoassay (EIA)/Enzyme-Linked Immunosorbent Assay (ELISA). Lequin RM, Clinical Chemistry. 2005; 51, 12:  2415-2418.

 

February is marked as “Heart Month” drawing attention to heart disease and its prevention. Cardiovascular diseases (CVD) are the leading cause of death globally with an estimated toll of 17.9 million people per year (1).

Unhealthy diet, physical inactivity, tobacco use are only some of the  behavioral risk factors associated with heart disease. Health conditions that increase the risk include obesity, elevated blood sugar levels, high blood pressure, high low-density lipoprotein (LDL) cholesterol, and oxidative stress.

Biomarkers of Oxidative Stress in Heart Disease

What is oxidative stress?

 

Oxidative stress is caused by an imbalance between the production of free radicals (reactive oxygen species- ROS) and antioxidant defenses (endogenous antioxidant capacity). ROSs are generated as metabolic by-products in cells that can be stimulated by a variety of agents including pollutants, heavy metals, tobacco, drugs and many other. It has been postulated that oxidative stress can be responsible in the onset and progression of several diseases (e.g. cancer, diabetes, metabolic disorders, atherosclerosis, and cardiovascular diseases) (2).

Oxidized LDL (oxLDL) and Anti-oxidized LDL Autoantibodies

Accumulation of oxidized-low density lipoproteins (oxLDL) in the blood vessels triggers the onset of atherosclerosis which is a key element of the development of cardiovascular disease (CVD). The term atherosclerosis is of Greek origin, meaning thickening of the intimal layer of arteries and accumulation of fat (3). The process of atherosclerosis is initiated by LDL particles that lie in the sub-endothelial space of arteries (4). Thus, atherosclerosis is a disease of the vascular intima as a result of lipid oxidation. In the later stages of disease progression, plaques build up that rupture resulting in thrombosis (blockage of blood flow). oxLDL induces inflammatory responses in macrophages and dendritic cells (5).

Autoantibodies against oxidatively modified LDL (anti-oxLDL Ab)  have been detected in patients with atherosclerosis as well as in healthy individuals (6). It has been suggested that they mirror the occurrence of oxidation processes taking place in vivo. Anti-oxLDL Ab can be measured in human blood samples with a conventional ELISA assay (6-8). Numerous studies have implied that autoantibodies to oxLDL may allow improved CV risk stratification (6).

How can oxidative status / oxidative stress be measured?

 

Oxidative stress can be measured indirectly by different techniques. 

Example of a BIOMEDICA ELISA kit

 

1.MEASUREMENT OF ANTI-OXIDIZED LDL AUTOANTIBODIES 

ELISA for the detection of Anti-oxidized LDL Autoantibodies (oLAB ) | BI-20032

• Sample type: serum
• Sample volume 50µl/well
• Incubation time: 1.5 h / 30 min / 15 min
• Detection range: 0 – 1200 mU / ml
• Sensitivity: 48 mU / ml
• Precision: In-between-run (n=5): ≤ 8 % CV, Within-run (n=8): ≤ 4 % CV
• Use: Research use only
• Widely cited in over 70 publications

 

2.  MEASUREMENT OF BIOLOGICAL PEROXIDES 

Oxidative Stress Test – OXYSTAT Assay | BI-5007

Quick and easy assay to measure total peroxides in biological fluids.

• Method: colorimetric assay, 96 wells
• Sample type: serum, plasma, biological fluids
• Sample volume 10µl/well
• Assay time: 15 min
• Detection range: 0 – 660 µmol/l
• Sensitivity: 7 µmol/l
• Use: Research use only
• Widely cited in over 50 publications

 

Results show a direct correlation between free radicals and circulating biological peroxides and thus allow the characterization of the oxidative status in biological samples.

Literature

1. WHO-international health topics-cardiovascular diseases.
2. Reactive oxygen species in the vasculature: molecular and cellular mechanisms. Taniyama Y, Griendling KK. Hypertension. 2003 Dec;42(6):1075-81. doi: 10.1161/01.HYP.0000100443.09293.4F. Epub 2003 Oct 27. PMID: 14581295.
3. Atherosclerosis: process, indicators, risk factors and new hopes. Rafieian-Kopaei M, Setorki M, Doudi M, Baradaran A, Nasri H. Int J Prev Med. 2014 Aug;5(8):927-46. PMID: 25489440; PMCID: PMC4258672.
4. Oxidized LDL and anti-oxidized LDL antibodies in atherosclerosis – Novel insights and future directions in diagnosis and therapy. Hartley A, Haskard D, Khamis R. Trends Cardiovasc Med. 2019 Jan;29(1):22-26. doi: 10.1016/j.tcm.2018.05.010. Epub 2018 Jun 4. PMID: 29934015.
5. How Oxidized Low-Density Lipoprotein Activates Inflammatory Responses. Rhoads JP, Major AS. Crit Rev Immunol. 2018;38(4):333-342. doi: 10.1615/CritRevImmunol.2018026483. PMID: 30806246; PMCID: PMC6527110.
6. Anti-Oxidized LDL Antibodies and Coronary Artery Disease: A Systematic Review. van den Berg VJ, Vroegindewey MM, Kardys I, Boersma E, Haskard D, Hartley A, Khamis R. Antioxidants (Basel). 2019 Oct 15;8(10):484. doi: 10.3390/antiox8100484. PMID: 31618991; PMCID: PMC6826549
7. Circulating oxidized low density lipoprotein, autoantibodies against them and homocysteine serum levels in diagnosis and estimation of severity of coronary artery disease. Faviou E, Vourli G, Nounopoulos C, Zachari A, Dionyssiou-Asteriou A. Free Radic Res. 2005 Apr;39(4):419-29. doi: 10.1080/10715760500072156. PMID: 16028367.
8. Antibodies to oxidized low-density lipoprotein in patients following coronary artery revascularization. Miller ER 3rd, Erlinger TP, Blumenthal RS, Margolis S, Allen JK. Coron Artery Dis. 2003 Apr;14(2):163-9. doi: 10.1097/00019501-200304000-00009. PMID: 12655280.

 

 

 

 

Biomedica´s Osteoprotegerin (OPG) and Fibroblast Growth Factor (FGF23) assays were highlighted in the German Chronic Kidney Disease (CKD) cohort study to investigate the prognostic value of a total of nine mineral and bone disease biomarkers (MBD) (1). Brief, 4246 participants were enrolled and baseline serum samples of the MBD markers were prospectively observed for a median time of 6.5 years. The markers were compared to study the association of each biomarker with adverse cardiovascular (CV) outcomes and mortality. Of the nine biomarkers examined, Osteoprotegerin (OPG) best identified CKD patients who had the highest risk. The findings mirror the link between bone metabolism and CV disease in CKD patients and may help clinicians to identify patients who are at high risk for CV disease (1).

OPG a marker of cardiovascular mortality in CKD

The Kidney, Bone, and Heart Connection

 Chronic Kidney Disease (CKD), is a condition involving a gradual loss of kidney function. The kidneys lose their ability to filter toxins and extra fluid from the blood. There is no cure for this progressive disease and it is associated with a high morbidity and mortality rate that occurs especially in people with diabetes and hypertension (2). Worldwide, it is estimated that CKD is present in one out of ten adults (3).

Bone Disorders in Chronic Kidney Disease

 Other important roles of the kidney include maintaining bone health and balancing important minerals in the body. A majority of patients  with CKD have an elevated risk of developing disturbances of bone and mineral metabolism that lead to bone lesions (4). Thus, mineral an bone disorders (MBD) with biochemical and hormonal alterations are part of the complications associated with the progression of CKD (5).

 

Cardiovascular Events in Chronic Kidney Disease (CKD)

 Mineral and bone disorder in CKD can not only affect the bones but also other organs including the heart and blood vessels. It is well recognized that both atherosclerosis and arteriosclerosis are accelerated in patients with kidney failure (6). The high cardiovascular morbidity and mortality in CKD patients is mainly caused by progressive vascular calcification.

Osteoprotegerin (OPG) a Cardiovascular Risk Factor in CKD

 Osteoprotegerin (OPG) is a protein that regulates bone mass by inhibiting bone resorption. Apart from its traditional role in bone, OPG plays an important role in vascular disease and calcification as it interacts with endothelial and smooth muscle cells (7).

In a large prospective, population based study, severity, initiation, and progression of atherosclerosis was assessed in carotid arteries (8). The researchers demonstrated that OPG was an independent risk factor for the progression of atherosclerosis and onset of cardiovascular disease (CV) (8).

Serum OPG has also been shown to be a predictor of progression of atherosclerosis and coronary calcification in hemodialysis patients (9) and OPG serum levels increase with CKD disease progression and are associated with mortality in these patients (10). Moreover, high OPG levels are also associated with CV events in the general population as demonstrated in a large community based cohort (10). These data strongly reinforce OPG as marker for CV disease risk factor burden and predictor of CVD and mortality in the community (11).

 

Finally, the data from the recent German Chronic Kidney Disease cohort study investigating the prognostic value of nine mineral and bone disease associated biomarkers (MBD) showed that only serum OPG levels consistently identified the CKD patients who had the highest risk for adverse CV outcomes and mortality (1).

Osteoprotegerin (OPG) and Fibroblast Growth Factor (FGF23) can reliably be measured with a conventional ELISA assay

 Biomedica OPG ELISA (cat. no. BI-20403)

• Method: Sandwich ELISA, HRP/TMB, 12×8-well detachable strips
• Sample: Serum, plasma (EDTA, citrate, heparin)
• Sample volume: 20μl / well
• Detection range: 1.25-20 pmol/L (= 25 – 400 pg/mL)
• Sensitivity: 0.07 pmol/L (= 1.4 pg/mL)
• Incubation time: 4 h + 1 h + 30 min (room temperature)
• Precision: In-between-run (n=12): ≤ 5 % CV: Within-run (n=5): ≤ 3 % CV
• Widely cited in over 250 publications!

 

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

• Method: Sandwich ELISA, HRP/TMB, 12×8-well detachable strips
• Sample: Serum, plasma (EDTA, citrate, heparin)
• Sample volume: 50μl / well
• Detection range: 0.2 – 20 pmol/mL (= 1.54 – 150.4 pg/mL)
• Sensitivity: 0.08 pmol/L (= 0.6 pg/mL)
• Incubation time: 20-24 h  + 1 h+ 30 min (room temperature)
• Precision: In-between (n=10): ≤ 10 % CV; Within-run (n=6): ≤ 12 % CV

 

Publications

1. Association of mineral and bone biomarkers with adverse cardiovascular outcomes and mortality in the German Chronic Kidney Disease (GCKD) cohort. Reimer KC, Nadal J, Meiselbach H, Schmid M, Schultheiss UT, Kotsis F, Stockmann H, Friedrich N, Nauck M, Krane V, Eckardt KU, Schneider MP, Kramann R, Floege J, Saritas T; GCKD study investigators. Bone Res. 2023 Oct 20;11(1):52. doi: 10.1038/s41413-023-00291-8. PMID: 37857629; PMCID: PMC10587182.
2. Chronic kidney disease. Kalantar-Zadeh K, Jafar TH, Nitsch D, Neuen BL, Perkovic V. Lancet. 2021 Aug 28;398(10302):786-802. doi: 10.1016/S0140-6736(21)00519-5. Epub 2021 Jun 24. PMID: 34175022.
3. 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. 20:100438. doi: 10.1016/j.lanepe.2022.100438. PMID: 36090671; PMCID: PMC9459126.
4. Chronic Kidney Disease-Mineral and Bone Disorder (CKD-MBD): Current Perspectives. Waziri B, Duarte R, Naicker S. Int J Nephrol Renovasc Dis. 2019 Dec 24;12:263-276. doi: 10.2147/IJNRD.S191156. PMID: 31920363; PMCID: PMC6935280.
5. Mineral Bone Disorders in Kidney Disease Patients: The Ever-Current Topic. Hu L, Napoletano A, Provenzano M, Garofalo C, Bini C, Comai G, La Manna G. Int J Mol Sci. 2022 Oct 13;23(20):12223. doi: 10.3390/ijms232012223. PMID: 36293076; PMCID: PMC9603742
6. Role of Chronic Kidney Disease (CKD)-Mineral and Bone Disorder (MBD) in the Pathogenesis of Cardiovascular Disease in CKD. Yamada S, Nakano T J Atheroscler Thromb. 2023 Aug 1;30(8):835-850. doi: 10.5551/jat.RV22006. Epub 2023 May 30. PMID: 37258233; PMCID: PMC10406631.
7. Role of crosstalk between endothelial cells and smooth muscle cells in vascular calcification in chronic kidney disease. Zhang YX, Tang RN, Wang LT, Liu BC. Cell Prolif. 2021. 54(3):e12980. doi: 10.1111/cpr.12980. Epub 2021 Jan 27. PMID: 33502070; PMCID: PMC7941222.
8. Osteoprotegerin is a risk factor for progressive atherosclerosis and cardiovascular disease. Kiechl S, Schett G, Wenning G, Redlich K, Oberhollenzer M, Mayr A, Santer P, Smolen J, Poewe W, Willeit J. Circulation. .109(18):2175-80. doi: 10.1161/01.CIR.0000127957.43874.BB. Epub 2004 Apr 26. PMID: 15117849.
9. Serum osteoprotegerin is a predictor of progression of atherosclerosis and coronary calcification in hemodialysis patients. Kurnatowska I, Grzelak P, Kaczmarska M, Stefańczyk L, Nowicki M. Nephron Clin Pract. 2011;117(4):c297-304. doi: 10.1159/000321169. Epub 2010 Sep 22. PMID: 20861651.
10. Circulating Osteoprotegerin in Chronic Kidney Disease and All-Cause Mortality. Kamińska J, Stopiński M, Mucha K, Pac M, Gołębiowski M, Niewczas MA, Pączek L, Foroncewicz B. Int J Gen Med. 2021 Jun 9;14:2413-2420. doi: 10.2147/IJGM.S302251. PMID: 34135625; PMCID: PMC8200134.
11. Biomarkers of the osteoprotegerin pathway: clinical correlates, subclinical disease, incident cardiovascular disease, and mortality.  Lieb W, Gona P, Larson MG, Massaro JM, Lipinska I, Keaney JF Jr, Rong J, Corey D, Hoffmann U, Fox CS, Vasan RS, Benjamin EJ, O’Donnell CJ, Kathiresan S. Arterioscler Thromb Vasc Biol. 2010 Sep;30(9):1849-54. doi: 10.1161/ATVBAHA.109.199661. Epub 2010 May 6. PMID: 20448212; PMCID: PMC3039214.
12. Association of mineral and bone biomarkers with adverse cardiovascular outcomes and mortality in the German Chronic Kidney Disease (GCKD) cohort. Reimer KC, Nadal J, Meiselbach H, Schmid M, Schultheiss UT, Kotsis F, Stockmann H, Friedrich N, Nauck M, Krane V, Eckardt KU, Schneider MP, Kramann R, Floege J, Saritas T; GCKD study investigators. Bone Res. 2023 Oct 20;11(1):52. doi: 10.1038/s41413-023-00291-8. PMID: 37857629; PMCID: PMC10587182.

 

 

 

 

Duchenne muscular dystrophy (DMD) is a hereditary neuromuscular disease that leads to progressive muscle fiber degeneration and weakness. There is no cure for this disease and current therapy consists on treatment with glycocorticoids (GC). GC therapy is linked to risk of bone loss and increased fracture risk. Despite their adverse effects GC remain the standard care to slow down disease progression (2).

Steroid therapy and fracture prevention in Duchenne Muscular Dystrophy

This recent study explored factors that are associated with incident fracture risk in glucocorticoid (GC)-treated patients with Duchenne muscular dystrophy (DMD): Risk Factors Associated with Incident Vertebral Fractures in Steroid-treated Males with Duchenne Muscular Dystrophy. Brief, vertical fractures (VF) were prospectively evaluated in 38 males with Duchenne muscular dystrophy at study baseline and 12 months . The authors concluded the following: ” The observation that ≥ 1 prevalent VF and/or non-VF were the strongest predictors of incident VFs at 12 months supports the need for prevention of first fractures in this high-risk setting. Bone age delay, a marker of GC exposure, may assist in the prioritization of patients in efforts to prevent first fractures.”

Steroid therapy and fracture prevention in Duchenne Muscular Dystrophy – The Biomedica IL-6 and Sclerostin ELISA were highlighted in this study.

High Sensitivity IL-6 ELISA  (cat. no. BI-IL6) – measurable values in serum and plasma samples.

• Format: 12×8 wells
• Sensitivity: 0.28 pg/ml
• Dynamic Range: 0-200 pg/ml
• Assay Time: 4 hours 30 minutes
• Sample Type: Serum, Plasma, Cell Culture Supernatants, Urine
• Sample Volume: 100 µl
• Alternative Name: Interleukin 6
• Kit includes 7 pre-diluted calibrators and 2 controls

 

Sclerostin ELISA (cat. no. BI-20492) 

• Format: 12×8 wells
• Sensitivity: 3.2 pmol/l (= 72 pg/ml)
• Dynamic Range: 0 to 240 pmol/l (=0-5400 pg/ml)
• Assay Time: 18-25h (overnight incubation) / 1 h /30 min
• Sample Type: Serum, Plasma, Cell Culture Supernatants, Urine
• Sample Volume: 20 µl
• Alternative Name: SOST
• Kit includes 6 pre-diluted calibrators and 1 control

 

Are you planning a study? Contact us to learn about our promotions info@bmgrp.com

Literature

1.Risk Factors Associated with Incident Vertebral Fractures in Steroid-treated Males with Duchenne Muscular Dystrophy. Phung K, McAdam L, Ma J, McMillan HJ, Jackowski S, Scharke M, Matzinger MA, Shenouda N, Koujok K, Jaremko JL, Wilson N, Walker S, Hartigan C, Khan N, Page M, Robinson ME, Saleh DS, Smit K, Rauch F, Siminoski K, Ward LM.J Clin Endocrinol Metab. 2023 Aug 23:dgad435. doi: 10.1210/clinem/dgad435. Epub ahead of print. PMID: 37610420.

Abstract

Purpose: Prevention of fractures is an unmet need in glucocorticoid (GC)-treated Duchenne muscular dystrophy. This study explored factors associated with incident vertebral fractures (VFs) to inform future fracture prevention efforts. Methods: VFs were evaluated prospectively at study baseline and 12 months on lateral spine radiographs in participants aged 4 to 25 years with Duchenne muscular dystrophy. Clinical factors were analyzed for their association with the change in Spinal Deformity Index (sum of the Genant-defined VF grades from T4 to L4) between baseline and 12 months. Results: Thirty-eight males were evaluated (mean ± SD age at baseline 11.0 ± 3.6 years; mean ± SD GC duration at baseline 4.1 ± 3.1 years; 74% ambulatory). Nine of 38 participants (24%) had 17 incident VFs, of which 3/17 VFs (18%) were moderate/severe. Participants with 12-month incident VF had lower mean ± SD baseline lumbar spine areal bone mineral density Z-scores (-2.9 ± 1.0 vs -1.9 ± 1.1; P = .049) and lower total body less head areal bone mineral density Z-scores (-3.1 ± 1.2 vs -1.6 ± 1.7; P = .036). Multivariable linear regression showed that at least 1 VF at baseline (P < .001), a higher number of antecedent non-VF (P < .001), and greater bone age delay at baseline (P = .027) were significant predictors of an increase in the Spinal Deformity Index from baseline to 12 months. Conclusion: The observation that ≥ 1 prevalent VF and/or non-VF were the strongest predictors of incident VFs at 12 months supports the need for prevention of first fractures in this high-risk setting. Bone age delay, a marker of GC exposure, may assist in the prioritization of patients in efforts to prevent first fractures.

2. Emerging therapies for Duchenne muscular dystrophy. Markati T, Oskoui M, Farrar MA, Duong T, Goemans N, Servais L. Lancet Neurol. 2022 Sep;21(9):814-829. doi: 10.1016/S1474-4422(22)00125-9. Epub 2022 Jul 15. PMID: 35850122.

 

 

 

BIOMEDICA provides custom analytical testing service for universities, biotechnology and pharmaceutical industries. Our skilled laboratory team conducts sample measurements for the quantification of biomarkers in various sample matrices. We deliver quality and reliable results tailored to your precise specifications. Our primary objective is to meet your specific goals! BIOMEDICA has a long track record in successfully conducting numerous research projects which have also resulted in publications in top journals.

BIOMEDICA Analytical Testing Service

ELISA ASSAYs for Biomarkers  DEVELOPED AND MANUFACTURED BY BIOMEDICA

We measure biomarkers using our propriety ELISA kits for the quantification of  analytes in serum, plasma, urine and cell culture supernatant. Check out our product portfolio 

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NEXT-GENERATION SEQUENCING & RT-QPCR MICRORNA SERVICES

We provide a broad range of high-quality RNA services including RNA extraction, NGS and RT-qPCR and the custom analysis of microRNA signatures.

• Click here to learn more about the workflow

 

Related literature

Enzyme-Linked Immunosorbent Assay (ELISA). Tabatabaei MS, Ahmed M. Methods Mol Biol. 2022;2508:115-134. doi: 10.1007/978-1-0716-23763_10. PMID: 35737237.

Abstract

Enzyme-linked immunosorbent assay (ELISA) is one of the most specific and straightforward assays for detecting biomolecules in research and clinics. With advances in analytical methods, ELISA assay has been constantly optimized to improve its sensitivity, and different types of ELISA are now available to detect various biomarkers. This chapter provides an overall summary of the basic principle of ELISA, discusses different components of ELISA assay, and clearly outline protocols for different types of ELISA assays, including direct, indirect, sandwich, competitive, and nanoparticle-based ELISA.

Circulating miRNAs in bone health and disease. Grillari J, Mäkitie RE, Kocijan R, Haschka J, Vázquez DC, Semmelrock E, Hackl M. Bone. 2021 Apr;145:115787. doi: 10.1016/j.bone.2020.115787. Epub 2020 Dec 8. PMID: 33301964.

Abstract

microRNAs have evolved as important regulators of multiple biological pathways essential for bone homeostasis, and microRNA research has furthered our understanding of the mechanisms underlying bone health and disease. This knowledge, together with the finding that active or passive release of microRNAs from cells into the extracellular space enables minimal-invasive detection in biofluids (circulating miRNAs), motivated researchers to explore microRNAs as biomarkers in several pathologic conditions, including bone diseases. Thus, exploratory studies in cohorts representing different types of bone diseases have been performed. In this review, we first summarize important molecular basics of microRNA function and release and provide recommendations for best (pre-)analytical practices and documentation standards for circulating microRNA research required for generating high quality data and ensuring reproducibility of results. Secondly, we review how the genesis of bone-derived circulating microRNAs via release from osteoblasts and osteoclasts could contribute to the communication between these cells. Lastly, we summarize evidence from clinical research studies that have investigated the clinical utility of microRNAs as biomarkers in musculoskeletal disorders. While previous reviews have mainly focused on diagnosis of primary osteoporosis, we have also included studies exploring the utility of circulating microRNAs in monitoring anti-osteoporotic treatment and for diagnosis of other types of bone diseases, such as diabetic osteopathy, bone degradation in inflammatory diseases, and monogenetic bone diseases.

Preeclampsia is characterized by persistent high blood pressure and signs of liver or kidney damage that can affect pregnant women usually in the second half of pregnancy. It can also occur in women after the baby is delivered, mostly within 48 hours. Preelampsia occurs in around 2-8% of all pregnancies worldwide and is considered as a major cause of maternal and fetal mortality (1,2). It is responsible for over 70 000 maternal deaths and 500 000 fetal deaths worldwide every year (2). The cause of preeclampsia is still under debate and the clinical symptoms appear late during pregnancy (3).  Early diagnosis is essential to prevent morbidity and mortality that is associated with preeclampsia (4).

Endostatin a biomarker for early prediction of preeclampsia  

Endostatin is one of the most potent inhibitors of angiogenesis as it specifically inhibits the proliferation and migration of endothelial cells. Endostatin is a degradation product of collagen XVIII, a protein of the extracellular matrix (ECM) which is expressed in the basement membranes. Endostatin is a stable analyte that can reliably be measured in human blood and urine samples.

Biochemical markers may be useful to predict preeclampsia early, prior to the onset of clinical signs. A recent study explored the potential use of the protein biomarker Endostatin for the early prediction of preeclampsia (5).

BIOMEDICA human ENDOSTATIN ELISA │BI-20742 

• RELIABLE – validated according to FDA/ICH/EMEA guidelines
• FLEXIBLE  – only ELISA that can measure human Endostatin in serum, plasma and urine
• OPTIMIZED for clinical samples – wide dynamic range
• TRUSTED – cited and sample values provided  
• EFFICIENT – 20µl sample/well; results in 4.5 hours

 

BIOMEDICA mouse/rat ENDOSTATIN ELISA │BI-20742MR

• Fully validated for pre-clinical research (click here)
• Control included

 

Example of a BIOMEDICA ELISA assay kit

Literature

1. Preeclampsia diagnosis and management. Best Pract Res Clin Anaesthesiol. Overton E, Tobes D, Lee A. 2022 May;36(1):107-121. doi: 10.1016/j.bpa.2022.02.003. Epub 2022 Feb 10. PMID: 35659948.
2. Preeclampsia: Pathophysiology, Challenges, and Perspectives. Rana S, Lemoine E, Granger JP, Karumanchi SA. Circ Res. 2019 Mar 29;124(7):1094-1112. doi: 10.1161/CIRCRESAHA.118.313276. Erratum in: Circ Res. 2020 Jan 3;126(1):e8. PMID: 30920918.
3. The placenta and preeclampsia: villain or victim? Melchiorre K, Giorgione V, Thilaganathan B. Am J Obstet Gynecol. 2022 Feb;226(2S):S954-S962. doi: 10.1016/j.ajog.2020.10.024. Epub 2021 Mar 24. PMID: 33771361.
4. Early Detection of Preeclampsia Using Circulating Small non-coding RNA. Yoffe L, Gilam A, Yaron O, Polsky A, Farberov L, Syngelaki A, Nicolaides K, Hod M, Shomron N. Sci Rep. 2018 Feb 21;8(1):3401. doi: 10.1038/s41598-018-21604-6. PMID: 29467498; PMCID: PMC5821867.
5. Endostatin and Cystatin C as Potential Biomarkers for Early Prediction of Preeclampsia. Alshannag F, Zaki RMM, Hemida E, ElBakry MMM, Noureldeen AFH. ACS Omega. 2023 Nov 1;8(45):42776-42786. doi: 10.1021/acsomega.3c05586. PMID: 38024766; PMCID: PMC10652833.

 

Citations

•  Endostatin Is an Independent Risk Factor of Graft Loss after Kidney Transplant. Chu C, Hasan AA, Gaballa MMS, Zeng S, Xiong Y, Elitok S, Krämer BK, Hocher B. Am J Nephrol. 2020;51(5):373-380. doi: 10.1159/000507824. Epub 2020 Apr 22. PMID: 32320989.
• Validation of an enzyme-linked immunosorbent assay (ELISA) for quantification of endostatin levels in mice as a biomarker of developing glomerulonephritis. Wallwitz J, Aigner P, Gadermaier E, Bauer E, Casanova E, Bauer A, Stoiber D. PLoS One. 2019 Aug 12;14(8):e0220935. doi: 10.1371/journal.pone.0220935. PMID: 31404120; PMCID: PMC6690585.
• Higher Parathyroid Hormone Level Is Associated With Increased Arterial Stiffness in Type 1 Diabetes. Zobel EH, Theilade S, von Scholten BJ, Persson F, Tarnow L, Lajer M, Hansen TW, Rossing P. Diabetes Care. 2017 Mar;40(3):e32-e33. doi: 10.2337/dc16-2428. Epub 2017 Jan 6. PMID: 28062524.
• Endostatin in chronic kidney disease: Associations with inflammation, vascular abnormalities, cardiovascular events and survival. Kanbay M, Afsar B, Siriopol D, Unal HU, Karaman M, Saglam M, Gezer M, Taş A, Eyileten T, Guler AK, Aydin İ, Oguz Y, Tarim K, Covic A, Yilmaz MI. Eur J Intern Med. 2016 Sep;33:81-7. doi: 10.1016/j.ejim.2016.06.033. Epub 2016 Jul 7. PMID: 27394925.

 

 

In kidney transplantation recipients antibody mediated rejection (AMR) is a common complication that contributes to graft loss. C4d staining in kidney biopsies is widely used to identify the presence of complement activation in the small blood vessels of the kidney. Therefore C4d has been recognized as a marker for identifying AMR in kidney transplants (1).

C4d a biomarker of transplant rejection

 

C4d antibodies by Biomedica – for the identification of human complement split product C4d in paraffin and frozen sections as well as by flow cytometry.

Anti-C4d Antibody | BI-RC4D

• widely cited in over 90 citations
• for immunohistochemistry on paraffin embedded tissue and frozen sections
• use in kidney, heart, liver and other transplants

 

Anti-C4d Antibody (FITC) | BI-RC4D-FITC

• protocol for cell- or solid-phase bound C4 and C4d split product by flow cytometry
• for kidney, heart, liver and other transplants

 

The Biomedica C4d antibody was highlighted in the following publication: Response to treatment and long-term outcomes in kidney transplant recipients with acute T cell-mediated rejection. “Complement split product C4d staining was performed by immunochemical analysis of paraffin sections using polyclonal human anti‐C4d antibodies (Biomedica Gruppe, Vienna, Austria)”.

 

Literature

1. The Banff 2019 Kidney Meeting Report (I): Updates on and clarification of criteria for T cell- and antibody-mediated rejection. Loupy A, Haas M, Roufosse C, Naesens M, Adam B, Afrouzian M, Akalin E, Alachkar N, Bagnasco S, Becker JU, Cornell LD, Clahsen-van Groningen MC, Demetris AJ, Dragun D, Duong van Huyen JP, Farris AB, Fogo AB, Gibson IW, Glotz D, Gueguen J, Kikic Z, Kozakowski N, Kraus E, Lefaucheur C, Liapis H, Mannon RB, Montgomery RA, Nankivell BJ, Nickeleit V, Nickerson P, Rabant M, Racusen L, Randhawa P, Robin B, Rosales IA, Sapir-Pichhadze R, Schinstock CA, Seron D, Singh HK, Smith RN, Stegall MD, Zeevi A, Solez K, Colvin RB, Mengel M. Am J Transplant. 2020 Sep;20(9):2318-2331. doi: 10.1111/ajt.15898. Epub 2020 May 28. PMID: 32463180; PMCID: PMC7496245.

 

Bone Health & Osteoporosis – Biomarkers of Bone Regulation

Maintaining strong and healthy bones is essential for our well-being. Bone not only provides structural support for the body but also protects vital organs and serves as a provider for minerals such as calcium and phosphorus. Bone density and bone strength are key components of bone health.

Osteoporosis is a condition of weakened and fragile bones. It is the most common metabolic bone disease in the world (1) that can affect individuals of various ages, but is more commonly associated with aging. Both men and women can be affected, but postmenopausal women are at higher risk to develop Osteoporosis due to the decline of hormonal estrogen levels which plays a protective role in bone. Prevention of Osteoporosis includes a balanced diet and exercise.

Bone remodeling is a continuous process that is tightly regulated between bone resorption of old or damaged bone and the formation of new bone. Various hormones and factors are involved in bone metabolism. The bone cycle consists of different phases and markers of bone metabolism can be categorized as markers of bone formation, markers of bone resorption and markers of the regulation of  bone metabolism.

Bone Health & Osteoporosis – biomarkers of bone regulation

Protein biomarkers are often used in clinical research or clinical settings to assess bone health and to monitor the effectiveness of Osteoporosis treatments. Some of these biomarkers provide information about the regulatory processes involved in bone metabolism and turnover.

Monitoring these biomarkers can provide information on the overall health of bones. Some of these regulatory biomarkers include:

• Sclerostin (SOST) – is produced by osteocytes, bone cells embedded in the bone. Sclerostin is a bone specific Wnt pathway inhibitor, that negatively regulates bone formation, by promoting osteoclastogenesis and bone resorption (3). Elevated Sclerostin levels may indicate decreased bone formation.

 

• Dickkopf-1 (DKK-1) – is like Sclerostin an inhibitor of Wnt signaling, which is crucial for bone formation. Elevated serum DKK-1 promote bone resorption (4). DKK-1 levels may indicated suppressed bone formation.

 

• Receptor Activator of Nuclear Factor-Kappa B Ligand (RANKL) – is a key regulator of osteoclast activation and formation. RANKL promotes bone resorption by activating the bone osteoclasts. RANKL is secreted by osteocytes and is the most important factor of osteoclast formation (5).  

 

• Osteoprotegerin (OPG) – is the decoy receptor for RANKL. OPG regulates bone resorption (6). Changes in the OPG / RANKL ratio can affect bone remodeling.

 

• Fibroblast growth factor (FGF23) – is a hormone that regulates phosphate homeostasis and vitamin D metabolism. Abnormal FGF23 levels are associated with disorders affecting bone health e.g. hypophosphatemic rickets (7). 

   

  These protein biomarkers can easily be measured in human blood samples with an ELISA assay

   

  BIOMEDICA´s Bone Marker ELISA kits

  

• Sclerostin (SOST) ELISA  (BI-20492)

  • Most referenced Sclerostin ELISA +290 citations
  • Low sample volume – 20µl / well
  • Validation following international guidelines

   

• Bioactive Sclerostin ELISA (BI-20472)

  • Targets the receptor binding region
  • Rigorously validated for clinical samples according to FDA/ICH/EMEA guidelines
  • Low sample volume – 20 µl of serum/plasma per well 

   

• Dickkopf-1 (DKK-1) ELISA  (BI-20413)

  • Widely cited +170 publications
  • Direct measurement
  • Validated following international guidelines

   

• Osteoprotegerin (OPG) ELISA (BI-20403)

  • most referenced human OPG ELISA in +250 citations
  • day test, ready to use color coded reagents
  • controls included

   

• Free soluble RANKL ELISA (BI-20462)

  • Highly sensitive  – measurable concentrations in healthy subjects
  • Only ELISA that measures free, uncomplexed soluble RANKL
  • Cited in over +300 citations 

   

• FGF23 c-terminal multi-matrix ELISA (BI-20702)

  • for serum and plasma samples
  • full validation 
  • cited in +50 publications

   

• FGF23 intact ELISA (BI-20700)

  • for serum and plasma samples
  • full validation 
  • one-step ELISA

  

  Literature

  1. The clinician’s guide to prevention and treatment of osteoporosis. LeBoff MS et al., Osteoporos Int. 2022 Oct;33(10):2049-2102. doi: 10.1007/s00198-021-05900-y.
  2. Role of Wnt signaling and sclerostin in bone and as therapeutic targets in skeletal disorders. Marini F et al., Osteoporos Int. 2023 Feb;34(2):213-238. doi: 10.1007/s00198-022-06523-7.
  3. Inflammation and Bone Metabolism in Rheumatoid Arthritis: Molecular Mechanisms of Joint Destruction and Pharmacological Treatments. Maeda K et al., Int J Mol Sci. 2022 Mar 6;23(5):2871. doi: 10.3390/ijms23052871.
  4. Osteocyte-Related Cytokines Regulate Osteoclast Formation and Bone Resorption. Kitaura H et al., Int J Mol Sci. 2020 Jul 21;21(14):5169. doi: 10.3390/ijms21145169.
  5. The Role of Osteoprotegerin in Vascular Calcification and Bone Metabolism: The Basis for Developing New Therapeutics. Rochette L et al., Calcif Tissue Int. 2019 Sep;105(3):239-251. doi: 10.1007/s00223-019-00573-6.
  6. FGF23-related hypophosphatemic rickets/osteomalacia: diagnosis and new treatment. Fukumoto S. J Mol Endocrinol. 2021 Feb;66(2):R57-R65. doi: 10.1530/JME-20-0089.

   

 

It is well recognized that exercise has beneficial effects on overall health (1). Skeletal muscle has been described as a “secretory organ” that produces cytokines in response to muscle contraction (2). Thus, muscle communicates with various organs and tissues e.g. adipose tissue, liver, pancreas, bones, and brain (1).

IL-6 in exercise and health

Interleukin-6 (IL-6) is a cytokine that is primarily known to play a key role in inflammation and the regulation of the immune response. However, IL-6 is also an important signaling molecule during exercise. Numerous studies have demonstrated that Il-6 is produced locally in working skeletal muscle (3, 4).  Exercise, in particular intense or prolonged physical activity increases circulating IL-6 levels contributing to the regulation of glucose and lipid metabolism (4). Additional effects of IL-6 are discussed in Exercise and health — emerging roles of IL-6.

IL-6 ELISA, cat. no. BI-IL6

• HIGHLY SENSITIVE – measurable values in serum and plasma samples
• EASY – ready to use calibrators and controls
• RELIABLE – full validation package

Related ELISA kits

VEGF ELISA, cat. no. BI-VEGF

• SMALL SAMPLE VOLUME – 10µl/well
• HIGH SENSITIVITY – measurable values in both serum and plasma 
• RELIABLE –  rigorously validated according to FDA/ICH/EMEA guidelines
• EASY –  ready to use calibrators &  controls included

Angiopoietin-2 ELISA, cat. no. BI-ANG2

• Immediate results – assay range is optimized for clinical samples
• Characterized antibodies – detects all Angiopoietin-2 isoforms 
• Rigorously validated –  according to FDA/ICH/EMEA guidelines

Mouse/Rat Angiopoietin-2 ELISA, cat. no. BI-ANG2MR

• Low sample volume- 5µl
• Optimized for preclinical samples
• Sample values provided
• Detects all Angiopoetin-2 isoforms

Literature

1. Exercise and health — emerging roles of IL-6. Ellingsgaard H, Hojman P, Pedersen BK. Current Opinion in Physiology. 2019. 10; 49-54.
2. The effect of exercise on cytokines: implications for musculoskeletal health: a narrative review. Docherty, S., Harley, R., McAuley, J.J. et al. 2022. BMC Sports Sci Med Rehabil 14, 5.
3. IL-6 signaling in acute exercise and chronic training: Potential consequences for health and athletic performance. Nash D, Hughes MG, Butcher L, Aicheler R, Smith P, Cullen T, Webb R. Scand J Med Sci Sports. 2023. 33(1):4-19.
4. Muscle-derived interleukin-6: possible biological effects. Pedersen BK, Steensberg A, Schjerling P. J Physiol. 2001. 15;536(Pt 2):329-37.

Sepsis-induced brain dysfunction, referred to as sepsis-associated encephalopathy (SAE), is a common complication of sepsis affecting over half of sepsis patients in critical care units. It is linked to increased rates of mortality, morbidity, and long-term cognitive disability (1). The evaluation of SAE is crucial as it may be present in early stages of sepsis even before the diagnostic criteria for sepsis may be met (2). Biomarkers that are associated with brain injury could provide insights into the pathophysiological mechanisms of SAE and may be useful in patients for whom clinical assessment is challenging. C-type natriuretic peptide (CNP) and its amino-terminal propeptide NT-proCNP are natriuretic peptides, expressed in the vascular endothelium of many organs including the brain (3). Inflammatory mediators like IL-1 β and TNF-α can induce the release of NT-proCNP, suggesting a connection between systemic inflammation and the secretion and the regulation of NT-proCNP (4).

NT-proCNP in sepsis-induced brain dysfunction

The specific role of NT-proCNP as a biomarker for the assessment of SAE has been studied by Ehler et al., investigating NT-proCNP plasma concentrations in patients diagnosed with septic encephalopathy. The study revealed that highly elevated NT-proCNP levels in the early phase of sepsis might serve as an indicator for predicting the onset of SAE as the disease progresses (5).

NT-proCNP can reliably be measured in biological fluids with an ELISA assay from BIOMEDICA.

NT-proCNP ELISA (cat. no. BI-20823) kit highlights 

o   EFFICIENT – 20µl/well sample volume

o   CONVENIENT – 7 ready to use calibrators and 2 controls included

o   QUALITY – full validation following international quality guidelines

o   TRUSTED – widely cited

– Protocols available for urine, cell culture and non-human samples

Example of a Biomedica ELISA kit

 

MEASUREMENT OF NT-proCNP in NON-HUMAN SAMPLES

The sequence homology of NT-proCNP is very conserved among different species. We therefore assessed if the assay which is validated for human samples can be used in rat, mouse and pig samples. Other species types that share a high homology to human NT-proCNP can likely be measured with this assay (e.g. monkey, cats, dogs).  Learn more (NT-proCNP ELISA validation file pages 15-17). 

Literature

1. Septic-Associated Encephalopathy: a Comprehensive Review. Mazeraud A, Righy C, Bouchereau E, Benghanem S, Bozza FA, Sharshar T. Neurotherapeutics. 2020 Apr;17(2):392-403. doi: 10.1007/s13311-020-00862-1. PMID: 32378026; PMCID: PMC7283452.
2. Sepsis Associated Encephalopathy. Chaudhry N, Duggal AK.Adv Med. 2014;2014:762320. doi: 10.1155/2014/762320. Epub 2014 Sep 30. PMID: 26556425; PMCID: PMC4590973.
3. Prognostic value of circulating amino-terminal pro-C-type natriuretic peptide in critically ill patients. Koch A, Voigt S, Sanson E, Dückers H, Horn A, Zimmermann HW, Trautwein C, Tacke F.Crit Care. 2011;15(1):R45. doi: 10.1186/cc10007. Epub 2011 Jan 31. PMID: 21281508; PMCID: PMC3221974.
4. Inflammatory cytokines and NT-proCNP in Parkinson’s disease patients. Koziorowski D, Tomasiuk R, Szlufik S, Friedman A.Cytokine. 2012 Dec;60(3):762-6. doi: 10.1016/j.cyto.2012.07.030. Epub 2012 Aug 19. PMID: 22910321.
5. Diagnostic value of NT-proCNP compared to NSE and S100B in cerebrospinal fluid and plasma of patients with sepsis-associated encephalopathy. Ehler J, Saller T, Wittstock M, Rommer PS, Chappell D, Zwissler B, Grossmann A, Richter G, Reuter DA, Nöldge-Schomburg G, Sauer M. Neurosci Lett. 2019 Jan 23;692:167-173. doi: 10.1016/j.neulet.2018.11.014. Epub 2018 Nov 10. PMID: 30423400.

 

Cardiovascular toxicity (CV) plays a significant role in the failure of drug development. The use of blood-based biomarkers provides a way to detect CV toxicity during the preclinical stage, enabling the prioritization of compounds that show a lower risk inducing such adverse effects.

Cardiac Safety Biomarkers

The natriuretic peptides NT-proANP and NT-proBNP are cardiac hormones that are secreted when the heart muscle stretches. These peptides can be measured in blood samples and are well-known preclinical safety biomarkers used throughout drug development e.g NT-proBNP and NT-proANP – markers of drug-induced hypertrophy in rats (1-3).

BIOMEDICA provides robust ELISA assays for the accurate measurement of the these cardiac hormones for both human and rodent samples.

BIOMEDICA´s NT-proANP ELISA kit (BI-20892) has been independently validated in preclinical toxicology testing in rats:

• Cross-laboratory analytical validation of the cardiac biomarker NT-proANP in rat. 

 

In brief, the assay was evaluated by the Predictive Safety Testing Consortium, Cardiac Hypertrophy Working Group (PSTC—CHWG) in a cross-laboratory (5 laboratories) analytical assessment for use with rat samples. In summary, the assay has proven to be robust and technically adequate for the detection of NT-proANP serum levels in SD rats.

NT-proANP ELISA assay (cat. no. BI-20892)  

• 10 µl / well, serum or plasma
• widely cited as cardiovascular safety biomarker in rats
• for use in human and non-human samples (high cross-reactivity between species)

 

Rat NT-pro BNP ELISA assay (cat. no. BI-1204R) NEW

contact us for your special evaluation discount  info@bmgrp.com

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

 

Example of a Biomedica ELISA kit

 

PRODUCT INFORMATION

NT-proANP ELISA 

• Product code: BI-20892
• Method: ELISA
• Time to result: 3.5 hours
• Sample types: Serum, plasma, urine, cell culture supernatant (human, rat, mouse samples)
• Sample volume: 10 µl/well
• Sensitivity LOD: 0.05 nmol/l (= 0.64 ng/ml)
• Standard curve range: 0 – 10 nmol/l (= 0 – 127 ng/ml)
• Specificity: Endogenous and recombinant human NT-proANP (equivalent to proANP 1-98). Very high sequence homology between human and rodent (rat, mouse, and other species e.g. rabbit).
• NT-proANP ELISA assay is suitable for rat and mouse samples
• Instructions for use: click here

 

Rat NT-proBNP ELISA 

• Product code: BI-1204R
• Method: ELISA
• Time to result: 3.5 hours
• Sample types: rat serum and plasma
• Sample volume: 10 µl/well
• Sensitivity LOD: 21 pg/ml
• Standard curve range: 0 – 3200 pg/ml
• Specificity: Endogenous and recombinant rat NT-proBNP
• Instructions for use click here

 

Also available:

NT-proBNP human ELISA (cat. no. SK-1204), CE-marked, kit includes 2 controls 

CARDIAC SAFETY BIOMARKER ASSAYS in PRECLINICAL TOXICOLOGY TESTING 

References/Citations/Related Literature

All Citations on  NT-proANP as a cardiovascular safety biomarker in rat and mouse samples , click for all references

1. 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.J Pharmacol Toxicol Methods. 2016; 77:58-65. doi: 10.1016/j.vascn.2015.10.002. PMID: 26516096.

2. 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 et al., Toxicol Pathol. 2017; 45(2):344-352.
3. Natriuretic Peptides as Cardiovascular Safety Biomarkers in Rats: Comparison With Blood Pressure, Heart Rate, and Heart Weight. Engle SK, Watson DE.Toxicol Sci. 2016; 149(2):458-72. doi: 10.1093/toxsci/kfv240. Epub 2015 Nov 25. PMID: 26609138.

 

 

Type 2 diabetes mellitus (T2DM) is characterized by a persistent state of elevated blood sugar levels and glucose intolerance, resulting from the body´s incomplete response to insulin, accompanied by an increase in insulin production and a subsequent insulin deficiency. Individuals suffering from T2DM have an increased risk of cardiovascular disease (CVD). High glucose levels, insulin resistance, and chronic inflammation, contribute to endothelial dysfunction (ED) and atherosclerosis (1). ED refers to an impairment of the endothelium, the inner lining of blood vessels, which play an important role in regulating vascular health.

Sclerostin is associated with endothelial dysfunction in patients with type 2 diabetes

Sclerostin is a protein known primarily for its role in bone metabolism. It has also been identified of being linked to endothelial dysfunction in individuals diagnosed with type 2 diabetes (2). Sclerostin is predominantly secreted by osteocytes, cells that are embedded in the bone. However, vascular endothelial cells have also been observed to produce sclerostin leading to the discovery of its significant anti-calcifying role (3).

Sclerostin is associated with endothelial dysfunction in patients with type 2 diabetes: In an investigation in individuals with T2DM, researchers measured endothelial dysfunction by digital thermal monitoring (2). This method is a valid and noninvasive technique to evaluate endothelial function using temperature change on finger as a surrogate measure of the magnitude of vascular reactivity index (VRI) (4) . Serum Sclerostin levels were measured in the T2DM cohort with the Biomedica ELISA.  The prospective cross-sectional study revealed that serum sclerostin levels are positively associated with endothelial dysfunction measured in patients with T2DM.

A previous cross-sectional study in patients with T2DM, with/without cardiovascular disease, determined Sclerostin levels and its expression by RT-qPCR and immunohistochemistry in calcified and non-calcified artery of the lower limb from T2D. Serum Sclerostin was measured with an ELISA from Biomedica. Moreover, in vitro experiments were performed in vascular smooth muscle cells under calcifying conditions investigating the cardioprotective function of Sclerostin (5). The study provided evidence that supports the protective function of Sclerostin in the development of vascular calcification. The findings suggest that Sclerostin could potentially reduce the susceptibility to atherosclerosis by decreasing atherosclerotic plaque formation and underscore the significance of the bone-vascular axis when developing therapeutic strategies for treating impaired bone metabolism or vascular diseases (5).

Features and Benefits when measuring Sclerostin with the Biomedica ELISA kits

Sclerostin ELISA, #BI-20492

• TRUSTED – most referenced Sclerostin ELISA (+280 citations)
• LOW sample volume – 20µl sample /well
• For SERUM & PLASMA samples
• RELIABLE – fully validated  following international quality guidelines

 

 Bioactive Sclerostin ELISA, BI-20472

• CHARACTERIZED ANTIBODIES – targeting the receptor binding region
• EXTENSIVELY validated for clinical samples
• LOW sample volume – 20µl sample /well 

 

Literature

1. Endothelial Cell Dysfunction and the Pathobiology of Atherosclerosis. Garcia-de Los Ríos C, Medina-Casado M, Díaz-Chamorro A, Sierras-Jiménez M, Lardelli-Claret P, Sclerostin as a biomarker of cardiovascular risk in women with systemic lupus erythematosus. Cáliz-Cáliz R, Sabio JM. Sci Rep. 2022 Dec 14;12(1):21621. doi: 10.1038/s41598-022-25651-y. PMID: 36517533; PMCID: PMC9749620. Gimbrone MA Jr, García-Cardeña G. Circ Res. 2016 Feb 19;118(4):620-36. doi: 10.1161/CIRCRESAHA.115.306301. PMID: 26892962; PMCID: PMC4762052.
2. Serum sclerostin level is positively associated with endothelial dysfunction measured by digital thermal monitoring in patients with type 2 diabetes: A prospective cross-sectional study. Hsu BG, Wu DA, Yang HY, Chen MC.Medicine (Baltimore). 2023 Sep 8;102(36):e34649. doi: 10.1097/MD.0000000000034649. PMID: 37682176; PMCID: PMC10489308.
3. Sclerostin Protects Against Vascular Calcification Development in Mice. De Maré A, Opdebeeck B, Neven E, D’Haese PC, Verhulst A J Bone Miner Res. 2022 Apr;37(4):687-699. doi: 10.1002/jbmr.4503. Epub 2022 Feb 15. PMID: 35038187; PMCID: PMC9303214.
4. Digital thermal monitoring techniques to assess vascular reactivity following finger and brachial occlusions. Heath M, Gourley D, Naghavi M, Klies S, Tanaka H. J Clin Hypertens (Greenwich). 2020 Dec 7;23(1):122–7. doi: 10.1111/jch.14115. PMCID: PMC8030097.
5. Cardioprotective function of sclerostin by reducing calcium deposition, proliferation, and apoptosis in human vascular smooth muscle cells. González-Salvatierra S, García-Fontana C, Lacal J, Andújar-Vera F, Martínez-Heredia L, Sanabria-de la Torre R, Ferrer-Millán M, Moratalla-Aranda E, Muñoz-Torres M, García-Fontana B. Cardiovasc Diabetol. 2023 Nov 2;22(1):301. doi: 10.1186/s12933-023-02043-8. PMID: 37919715; PMCID: PMC10623848.

November is recognized as Diabetes Awareness Month, raising attention about diabetes and the importance of prevention and management of the disease that impacts millions of people worldwide.

Diabetes and Heart Disease – Risk stratification with NT-proBNP

Diabetes and heart disease are closely linked and patients suffering from diabetes have a 2- to 4 fold increased risk for developing cardiovascular diseases (CVD)  and concomitant heart failure (HF). Furthermore, HF patients with diabetes mellitus (DM) have a worse prognosis that those patients without DM (1, 2). In individuals with type 2 diabetes, high blood sugar levels and insulin resistance can lead to inflammation and atherosclerosis, a disease where arteries narrow, increasing the risk of CVD. Altered blood sugar levels are often associated with other CV risk factors, such as hypertension and obesity (3).

Diabetes and Heart Disease – Risk stratification with NT-proBNP 

Identifying patients who are at risk in developing HF remains a clinical challenge. Assessing the risk of HF development in patients with diabetes is important and the use of biomarkers could be beneficial (4). NT-proBNP, a protein that is specifically secreted by cardiac cells, is the “gold standard” biomarker for heart failure (5, 6). In patients with type 2 diabetes, NT-proBNP has been shown to predict death and cardiovascular events (7, 8). Moreover, a recent study investigated the relationships of NT-proBNP with the risk of developing diabetes and diabetes-related complication in initially healthy individuals (9). NT-pro BNP levels were measured in blood samples in a large cohort (European Prospecive Investigation Into Cancer and Nutrition) of over 27,000 individuals. The study demonstrated that NT-proBNP concentrations were inversely associated with the risk of type 2 diabetes in healthy individuals. Higher NT-proBNP levels are related to a higher risk of vascular disease in people who develop the disease. The authors conclude that NT-proBNP may be a promising biomarker to identify asymptomatic individuals who are at high risk developing diabetes independent of potential confounders.

NT-proBNP can be reliably measured in SERUM & PLASMA with a conventional ELISA assay.

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

• RELIABLE – CE marked  –  for IVD use in the EU
• FLEXIBLE & EASY–  can be run in every lab
• CONVENIENT – Two controls included
• Assay independently validated in saliva samples
• Proficiency tested in accredited laboratory program Proficiency Testing Certificate 

 

Example of a BIOMEDICA ELISA assay test kit.

NT-proBNP ELISA (#SK-1204) click here for more information

Related products

NT-proANP ELISA (cat. no. BI-20892) – for human and rodent samples (10µl sample volume)

Rat NT-proBNP ELISA (cat. no. BI-1204R)  (10µl sample volume)

Literature

1. Epidemiology, Pathophysiology, Diagnosis and Treatment of Heart Failure in Diabetes. Park JJ. Diabetes Metab J. 2021 Mar;45(2):146-157. doi: 10.4093/dmj.2020.0282. Epub 2021 Mar 25. Erratum in: Diabetes Metab J. 2021 Sep;45(5):796. PMID: 33813813; PMCID: PMC8024162.
2. Diabetes as a risk factor for heart failure in women and men: a systematic review and meta-analysis of 47 cohorts including 12 million individuals. Ohkuma T, Komorita Y, Peters SAE, Woodward M. Diabetologia. 2019 Sep;62(9):1550-1560. doi: 10.1007/s00125-019-4926-x. Epub 2019 Jul 18. PMID: 31317230; PMCID: PMC6677875.
3. Diabetes leading to heart failure and heart failure leading to diabetes: epidemiological and clinical evidence. Palazzuoli A, Iacoviello M. Heart Fail Rev. 2023 May;28(3):585-596. doi: 10.1007/s10741-022-10238-6. Epub 2022 May 6. PMID: 35522391; PMCID: PMC10140137.
4. Assessing Cardiovascular Risk in Patients with Diabetes: An Update. Damaskos C, Garmpis N, Kollia P, Mitsiopoulos G, Barlampa D, Drosos A, Patsouras A, Gravvanis N, Antoniou V, Litos A, Diamantis E. Curr Cardiol Rev. 2020;16(4):266-274. doi: 10.2174/1573403X15666191111123622. PMID: 31713488; PMCID: PMC7903509.
5. NT-proBNP: The Gold Standard Biomarker in Heart Failure. McKie PM, Burnett JC Jr. J Am Coll Cardiol. 2016 Dec 6;68(22):2437-2439. doi: 10.1016/j.jacc.2016.10.001. PMID: 27908348.
6. 2022 AHA/ACC/HFSA Guideline for the Management of Heart Failure: Executive Summary: A Report of the American College of Cardiology/American Heart Association Joint Committee on Clinical Practice Guidelines. Heidenreich PA, Bozkurt B, Aguilar D, Allen LA, Byun JJ, Colvin MM, Deswal A, Drazner MH, Dunlay SM, Evers LR, Fang JC, Fedson SE, Fonarow GC, Hayek SS, Hernandez AF, Khazanie P, Kittleson MM, Lee CS, Link MS, Milano CA, Nnacheta LC, Sandhu AT, Stevenson LW, Vardeny O, Vest AR, Yancy CWCirculation. 2022 May 3;145(18):e876-e894. doi: 10.1161/CIR.0000000000001062. Epub 2022 Apr 1. PMID: 35363500.
7. NT-proBNP as a predictor of death and cardiovascular events in patients with type 2 diabetes. Malachias MVB, Wijkman MO, Bertoluci MC. Diabetol Metab Syndr. 2022 May 3;14(1):64. doi: 10.1186/s13098-022-00837-6. PMID: 35501909; PMCID: PMC9063067.
8. The role of NT-proBNP in the evaluation of diabetic patients with heart failure. Fringu FI, Sitar-Taut AV, Caloian B, Zdrenghea D, Comsa D, Gusetu G, Pop D. Acta Endocrinol (Buchar). 2020 Apr-Jun;16(2):183-191. doi: 10.4183/aeb.2020.183. PMID: 33029235; PMCID: PMC7535893.
9. Opposing Associations of NT-proBNP With Risks of Diabetes and Diabetes-Related Complications. Birukov A, Eichelmann F, Kuxhaus O, Polemiti E, Fritsche A, Wirth J, Boeing H, Weikert C, Schulze MB. Diabetes Care. 2020 Dec;43(12):2930-2937. doi: 10.2337/dc20-0553. Epub 2020 Aug 17. PMID: 32816995; PMCID: PMC7770272.