Analytical performance evaluation of sensitive and old generation reagent in routine practical use: estradiol experience

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Evaluation of the analytical performance of tests in medical laboratories is important. Total Error (TE) and sigma analysis have been used as a quantitative indicator of quality for many years. The aim of this study is to evaluate the analytical performance of Beckman Coulter Access Estradiol (E2) and Sensitive E2 reagents. Analytical performance of two reagents were evaluated with TE, six sigma and measurement uncertainty values. Two Beckman Coulter Unicel DxI-800 autoanalyzers (A1 and A2) included in the study. Quality control data between December 2017 and December 2019 were used. CLIA-2019 values were used for total allowable error (TEa) limits. Uncertainty values were calculated with ISO/TS 20914.
The median TE of the old generation and sensitive E2 reagent were 27.46% (between 13.49 and 48.88) and 11.16% (between 7.08 and 24.81), respectively (p < .005) The process sigma results were below 3 sigma in all months with the old reagent, whereas with the new reagents it was seen to be above 3 sigma in 11 of 12 months for both autoanalyzers. Uncertainty of old reagent is higher than new reagent. Imprecisions decrease as concentration increases with both reagents. The uncertainty values of low concentration levels are greater than high concentration levels. In conclusion, in both auto analyzers, Sensitive E2 reagent was found to have better performance than old reagent in terms of TE, process sigma and measurement uncertainty. We believe that the sensitive E2 reagent still needs further improvement for patients who have low E2 levels.

Prothymosin Alpha: A Novel Contributor to Estradiol Receptor Alpha-Mediated CD8 + T-Cell Pathogenic Responses and Recognition of Type 1 Collagen in Rheumatic Heart Valve Disease

Background: Rheumatic heart valve disease (RHVD) is a leading cause of cardiovascular death in low- and middle-income countries and affects predominantly women. The underlying mechanisms of chronic valvular damage remain unexplored and regulators of sex predisposition are unknown.
Methods: Proteomics analysis of human heart valves (nondiseased aortic valves, nondiseased mitral valves [NDMVs], valves from patients with rheumatic aortic valve disease, and valves from patients with rheumatic mitral valve disease; n=30) followed by system biology analysis identified ProTα (prothymosin alpha) as a protein associated with RHVD. Histology, multiparameter flow cytometry, and enzyme-linked immunosorbent assay confirmed the expression of ProTα. In vitro experiments using peripheral mononuclear cells and valvular interstitial cells were performed using multiparameter flow cytometry and quantitative polymerase chain reaction. In silico analysis of the RHVD and Streptococcus pyogenes proteomes were used to identify mimic epitopes.
Results: A comparison of NDMV and nondiseased aortic valve proteomes established the baseline differences between nondiseased aortic and mitral valves. Thirteen unique proteins were enriched in NDMVs. Comparison of NDMVs versus valves from patients with rheumatic mitral valve disease and nondiseased aortic valves versus valves from patients with rheumatic aortic valve disease identified 213 proteins enriched in rheumatic valves. The expression of the 13 NDMV-enriched proteins was evaluated across the 213 proteins enriched in diseased valves, resulting in the discovery of ProTα common to valves from patients with rheumatic mitral valve disease and valves from patients with rheumatic aortic valve disease. ProTα plasma levels were significantly higher in patients with RHVD than in healthy individuals. Immunoreactive ProTα colocalized with CD8+ T cells in RHVD. Expression of ProTα and estrogen receptor alpha correlated strongly in circulating CD8+ T cells from patients with RHVD. Recombinant ProTα induced expression of the lytic proteins perforin and granzyme B by CD8+ T cells as well as higher estrogen receptor alpha expression. In addition, recombinant ProTα increased human leukocyte antigen class I levels in valvular interstitial cells. Treatment of CD8+ T cells with specific estrogen receptor alpha antagonist reduced the cytotoxic potential promoted by ProTα. In silico analysis of RHVD and S pyogenes proteomes revealed molecular mimicry between human type 1 collagen epitope and bacterial collagen-like protein, which induced CD8+ T-cell activation in vitro.
Conclusions: ProTα-dependent CD8+ T-cell cytotoxicity was associated with estrogen receptor alpha activity, implicating ProTα as a potential regulator of sex predisposition in RHVD. ProTα facilitated recognition of type 1 collagen mimic epitopes by CD8+ T cells, suggesting mechanisms provoking autoimmunity.

Pharmacokinetic Interaction Between the MEK1/MEK2 Inhibitor Trametinib and Oral Contraceptives Containing Norethindrone and Ethinyl Estradiol in Female Patients With Solid Tumors

This phase 1 postapproval study assessed the effect of the mitogen-activated protein kinase kinase enzyme 1/enzyme 2 inhibitor trametinib (2 mg once daily, repeat dosing) on the pharmacokinetics of combined oral contraceptives (COCs) containing norethindrone (NE; 1 mg daily) and ethinyl estradiol (EE; 0.035 mg daily) in 19 female patients with solid tumors. Compared with NE/EE administered without trametinib, NE/EE administered with steady-state trametinib was associated with a clinically nonrelevant 20% increase in NE exposure (area under the curve [AUC]) and no effect on EE exposure (geometric mean ratio [geo-mean] of NE/EE + trametinib to NE/EE [90%CI]: NE AUC calculated to the end of a dosing interval at steady-state [AUCtau ] 1.20 [1.02-1.41]; NE AUC from time zero to the last measurable concentration sampling time [AUClast ] 1.2 [0.999-1.45]; EE AUCtau 1.06 [0.923-1.22]; EE AUClast 1.05 [0.883-1.25]).
Maximum serum concentration (Cmax ) of NE increased by 13% and Cmax of EE decreased by 8.5% when dosed with steady-state trametinib compared with COCs administered alone (geo-mean ratio [90%CI]: NE Cmax 1.13 [0.933-1.36]; EE Cmax 0.915 [0.803-1.04]). These results indicate that repeat-dose trametinib does not lower exposure to NE or EE and, hence, is unlikely to impact the contraceptive efficacy of COCs. The pharmacokinetic parameters of trametinib and its metabolite M5 were consistent with historic data of trametinib alone. Coadministration of trametinib and COCs was generally well tolerated in this study, with observed safety signals consistent with the known safety profile of trametinib and no new reported safety events. Overall, the findings indicate that hormonal COCs can be coadministered in female patients who receive trametinib monotherapy without compromising the contraceptive efficacy.

Estradiol
A8425-1000 ApexBio 1g 41.6 EUR

Estradiol
A8425-5.1 ApexBio 10 mM (in 1mL DMSO) 40 EUR

Estradiol
A8425-50 ApexBio 50 mg 157.2 EUR

Estradiol
317788 MedKoo Biosciences 500.0mg 195 EUR

Estradiol
20-abx185851 Abbexa
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Estradiol
DE2693 Demeditec Diagnostics 96 84 EUR

Estradiol
AT070 Unibiotest 1mg 1641.6 EUR

Estradiol
AG070 Unibiotest 1 mg 627.6 EUR

Estradiol
MBS122090-01mg MyBiosource 0.1mg 220 EUR

Estradiol
MBS122090-1mg MyBiosource 1mg 350 EUR

Estradiol
MBS122090-5x1mg MyBiosource 5x1mg 1490 EUR

Estradiol
MBS122091-01mg MyBiosource 0.1mg 220 EUR

Estradiol
MBS122091-1mg MyBiosource 1mg 350 EUR

Estradiol
MBS122091-5x1mg MyBiosource 5x1mg 1490 EUR

Estradiol
GP9683 Glentham Life Sciences 1g 304.24 EUR

Estradiol
GP9683-1 Glentham Life Sciences 1 27.8 EUR

Estradiol
GP9683-10 Glentham Life Sciences 10 172.4 EUR

Estradiol
GP9683-10G Glentham Life Sciences 10 g 244.8 EUR

Estradiol
GP9683-1G Glentham Life Sciences 1 g 69.6 EUR

Estradiol
GP9683-25 Glentham Life Sciences 25 332.2 EUR

Estradiol
GP9683-25G Glentham Life Sciences 25 g 438 EUR

Estradiol
GP9683-5 Glentham Life Sciences 5 98.8 EUR

Sucrose acetate isobutyrate (SAIB) as a delivery vehicle for estradiol and sulpiride: Evaluation of endocrine responses in geldings and ovarian response in seasonally anovulatory mares

Sulpiride in vegetable shortening (VS) stimulates prolactin in horses for up to 10 days. Although effective, a pharmaceutical grade vehicle is needed for clinical application of sulpiride in horses. Sucrose acetate isobutyrate (SAIB), a hydrophobic polymer, may be an alternative to VS. Four in vivo experiments assessed the efficacy of SAIB for delivery of sulpiride, estradiol cypionate (ECP), and estradiol benzoate (EB). The first three studies utilized geldings to compare prolactin and luteinizing hormone (LH) concentrations between sulpiride delivered in VS and SAIB, and ECP or EB delivered in SAIB. Sulpiride stimulated (P < .01) prolactin similarly between vehicles. Geldings pre-treated with EB had higher (P < .05) prolactin responses to sulpiride compared to ECP-treated geldings on days 5, 6 and 9.
Both estradiol-sulpiride treatments stimulated LH with no differences between ECP and EB. Experiment 3 compared a simultaneous injection of EB-sulpiride to a non-simultaneous injection (one day apart) of EB-sulpiride. Prolactin was stimulated (P < .05) in both treatment groups, but the response lasted 2 days longer in geldings treated a day apart. Plasma LH increased (P < .01) in both groups equally for 10 days. Experiment 4 applied simultaneous and non-simultaneous EB-sulpiride treatments to seasonally anovulatory mares to induce ovarian activity. Prolactin and LH were stimulated similarly between treatments; however, non-simultaneously treated mares tended (P = .07) to have an ovarian response earlier. In conclusion, SAIB was a suitable vehicle for administration of estradiol and sulpiride and could be an alternative to VS for sustained-release drug delivery.

EVALUATION OF EPIANDROSTERONE AS A LONG-TERM MARKER OF TESTOSTERONE USE

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  • Identification and evaluation of long-term markers is crucial in prolonging the detection window for anabolic steroid abuse in sport. Recently, sulfoconjugated epiandrosterone was identified as a potential long-term marker for the abuse of certain endogenous anabolic agents, including testosterone, which continues to be widely used as a performance enhancing agent in sport. To evaluate the applicability of epiandrosterone sulfate as a marker for testosterone use, administration studies were conducted with multiple modes of testosterone administration – transdermal, intramuscular, and subcutaneous.
  • A modified sample preparation method was used to collect both glucuronidated and sulfoconjugated analytes of interest. Carbon isotope ratio measurements from the administration studies are presented here. Epiandrosterone was less effective than the conventionally used target compounds for detection of the low dose application (transdermal gel). With intramuscular administration, epiandrosterone was more diagnostic than with transdermal administration, but it did not prolong the detection window more than the conventional target compounds.
  • With subcutaneous administration, the doses administered to the subjects were varied and the effect in the epiandrosterone values was dependent on the magnitude of the dose administered. Epiandrosterone does not appear to be a useful marker in the detection of low dose testosterone administration. It is responsive to higher dose administration, but it does not provide extension of the detection window relative to conventional target compounds.

Development and validation of a fast gas chromatography combustion isotope ratio mass spectrometry method for the detection of epiandrosterone sulfate in urine.

In doping control, to confirm the exogenous origin of exogenously administered anabolic androgenic steroids (AAS), a gas chromatography combustion isotope ratio mass spectrometry (GC-C-IRMS) analysis is performed. Recently published work suggests epiandrosterone sulfate (EpiAS) as a promising IRMS target compound for the detection of AAS, capable of prolonging the detection window. However, EpiAS is only excreted in urine in its sulfoconjugated form while all other IRMS target compounds are excreted glucuronidated, meaning that EpiAS cannot be incorporated in the existing IRMS methods.
A separate extensive sample preparation needs to be performed on this compound with a different hydrolysis and extraction procedure and a different liquid chromatography (LC) clean-up. The current work presents a new, fast and easy to implement EpiAS IRMS method. The approach was based on the direct GC analysis of non-hydrolysed EpiAS, making the solid phase extraction, hydrolysis and acetylation step redundant.
Sample preparation consisted of a simple liquid liquid extraction, followed by LC fraction collection. A population study was performed to check the compliance with the criteria drafted by the World Anti-Doping Agency (WADA). To verify the applicability of the developed approach, the method was applied to the samples of four administration studies (i.e., dehydroepiandrosterone (DHEA), testosterone gel (T gel), androstenedione (ADION) and intramuscular testosterone undecanoate. In contrast to previously published data, the strength of EpiAS as target compound and the prolongation of the detection window in comparison with the conventional IRMS target compounds was less pronounced.

Synthesis and bioevaluation of novel steroidal isatin conjugates derived from epiandrosterone/androsterone.

Steroids are classes of natural products widely distributed in nature, which have been demonstrated to exhibit broad biological functions, and have also attracted increasing interest from bioorganic and pharmaceutical researches. In order to develop novel chemical entities as potential cytotoxic agents, a series of steroidal isatin conjugations derived from epiandrosterone and androsterone were efficiently prepared and characterized, and all these obtained compounds were screened for their potential cytotoxic activities.
The preliminary bioassay indicated that most of the newly synthesized compounds exhibited good cytotoxic activities against human gastric cancer (SGC-7901), melanoma (A875), and hepatocellular liver carcinoma (HepG2) cell lines compared with 5-fluorouracil (5-FU), which might be considered as promising scaffold for further development of potential anticancer agents.

The quantitation of 7beta-hydroxy-epiandrosterone in the plasma and seminal plasma of men with different degrees of fertility.

7beta-hydroxy-epiandrosterone (7beta-OH-EpiA) is an endogenous androgen metabolite that has been shown to exert neuroprotective, anti-inflammatory and anti-estrogenic effects. However, to the best of our knowledge no information is available about this androgen steroid in relation to sperm quality. We analyzed 7beta-OH-EpiA in plasma and seminal plasma using a newly developed isotope dilution ultra-high performance liquid chromatography – mass spectrometry method. Validation met the requirements of FDA guidelines. Levels of 7beta-OH-EpiA were measured in 191 men with different degrees of infertility. One-way analysis of variance followed by multiple comparison and correlation analysis adjusted for age, BMI and abstinence time were performed to evaluate the relationships between this steroid and sperm quality.
Concentrations of 7beta-OH-EpiA in seminal plasma were significantly higher in severely infertile men in comparison with healthy men and slightly infertile men. The same trend was found when blood plasma was evaluated. Furthermore, plasma 7beta-OH-EpiA negatively correlated with sperm concentration (-0.215; p<0.01) and total count (-0.15; p<0.05). Seminal 7beta-OH-EpiA was negatively associated with motility (-0.26; p<0.01), progressively motile spermatozoa (-0.233; p<0.01) and nonprogressively motile spermatozoa (-0.188; p<0.05). 7beta-OH-EpiA is associated with lower sperm quality and deserves more research in that respect.

Epiandrosterone sulfate prolongs the detectability of testosterone, 4-androstenedione, and dihydrotestosterone misuse by means of carbon isotope ratio mass spectrometry.

In the course of investigations into the metabolism of testosterone (T) by means of deuterated T and hydrogen isotope ratio mass spectrometry, a pronounced influence of the oral administration of T on sulfoconjugated steroid metabolites was observed. Especially in case of epiandrosterone sulfate (EPIA_S), the contribution of exogenous T to the urinary metabolite was traceable up to 8 days after a single oral dose of 40 mg of T. These findings initiated follow-up studies on the capability of EPIA_S to extend the detection of T and T analogue misuse by carbon isotope ratio (CIR) mass spectrometry in sports drug testing. Excretion study urine samples obtained after transdermal application of T and after oral administration of 4-androstenedione, dihydrotestosterone, and EPIA were investigated regarding urinary concentrations and CIR.
With each administered steroid, EPIA_S was significantly depleted and prolonged the detectability when compared to routinely used steroidal target compounds by a factor of 2 to 5. In order to simplify the sample preparation procedure for sulfoconjugated compounds, enzymatic cleavage by Pseudomonas aeruginosa arylsulfatase was tested and implemented into CIR measurements for the first time. Further simplification was achieved by employing multidimensional gas chromatography to ensure the required peak purity for CIR determinations, instead of sample purification strategies using liquid chromatographic fractionation. Taking into account these results that demonstrate the unique and broad applicability of EPIA_S for the detection of illicit administrations of T or T-related steroids, careful consideration of how this steroid can be implemented into routine doping control analysis appears warranted.

An omics investigation into chronic widespread musculoskeletal pain reveals epiandrosterone sulfate as a potential biomarker.

Chronic widespread musculoskeletal pain (CWP) is common, having a population prevalence of 10%. This study aimed to define the biological basis of the CWP/body mass association by using a systems biology approach. Adult female twins (n = 2444) from the TwinsUK registry who had extensive clinical, anthropometric, and “omic” data were included. Nontargeted metabolomics screening including 324 metabolites was carried out for CWP and body composition using dual-energy X-ray absorptiometry. The biological basis of these associations was explored through a genome-wide association study and replicated in an independent population sample (Cooperative Health Research in the Region of Augsburg [KORA] study, n = 2483). A causal role for the genetic variants identified was sought in CWP using a Mendelian randomisation study design.
Fat mass/height2 was the body composition variable most strongly associated with CWP (TwinsUK: P = 2.4 × 10(-15) and KORA: P = 1.59 × 10(-10)). Of 324 metabolites examined, epiandrosterone sulfate (EAS) was highly associated with both CWP (P = 1.05 × 10(-09) in TwinsUK and P = 3.70 × 10(-06) in KORA) and fat mass/height2. Genome-wide association study of EAS identified imputed single nucleotide polymorphism rs1581492 at 7q22.1 to be strikingly associated with EAS levels (P ≤ 2.49 × 10(-78)), and this result was replicated in KORA (P = 2.12 × 10(-9)). Mendelian randomization by rs1581492 genotype showed that EAS is unlikely to be causally related to CWP. Using an agnostic omics approach to focus on the association of CWP with body mass index, we have confirmed a steroid hormone association and identified a genetic variant upstream of the CYP genes, which likely controls this response. This study suggests that steroid hormone abnormalities result from pain rather than causing it, and EAS may provide a biomarker that identifies subgroups at risk of CWP.

Epiandrosterone
B1505-1000 ApexBio 1 g 40 EUR

Epiandrosterone
B1505-5.1 ApexBio 10 mM (in 1mL DMSO) 135.6 EUR

Epiandrosterone
B1505-S ApexBio Evaluation Sample 97.2 EUR

Epiandrosterone
HY-I0352 MedChemExpress 500mg 129.6 EUR

Epiandrosterone
GP6146 Glentham Life Sciences 1g 29.04 EUR

Epiandrosterone
GP6146-1 Glentham Life Sciences 1 31.7 EUR

Epiandrosterone
GP6146-1G Glentham Life Sciences 1 g 74.4 EUR

Epiandrosterone
E578000 Toronto Research Chemicals 1g 164 EUR

Epiandrosterone
MBS3604607-100mg MyBiosource 100mg 200 EUR

Epiandrosterone
MBS3604607-5x100mg MyBiosource 5x100mg 580 EUR

Epiandrosterone
MBS340129-10mg MyBiosource 10mg 3105 EUR

Epiandrosterone
MBS340129-1mg MyBiosource 1mg 800 EUR

Epiandrosterone
MBS340129-5x10mg MyBiosource 5x10mg 13805 EUR

Epiandrosterone
MBS342094-05mL MyBiosource 0.5mL 790 EUR

Epiandrosterone
MBS342094-5x05mL MyBiosource 5x0.5mL 3385 EUR

Epiandrosterone
TRC-E578000-1G Toronto Research Chemicals 1 g Ask for price

Epiandrosterone
TRC-E578000-250MG Toronto Research Chemicals 250 mg Ask for price

Epiandrosterone
TRC-E578000-5G Toronto Research Chemicals 5 g Ask for price

Epiandrosterone (HRP)
abx284059-100g Abbexa 100 µg Ask for price

Epiandrosterone (HRP)
abx284059-20g Abbexa 20 µg 862.5 EUR

Epiandrosterone (HRP)
abx284059-50g Abbexa 50 µg Ask for price

Epiandrosterone-d5
E578002 Toronto Research Chemicals 10mg 201 EUR

Epiandrosterone-d5
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Epiandrosterone-d5
TRC-E578002-10MG Toronto Research Chemicals 10 mg Ask for price

Epiandrosterone-HRP
MBS5304382-05mL MyBiosource 0.5mL 1485 EUR

Epiandrosterone-HRP
MBS5304382-5x05mL MyBiosource 5x0.5mL 6535 EUR

Epiandrosterone-HRP
80-1155 Fitzgerald Industries International 500 ul Ask for price

Epiandrosterone 17 (HRP)
MBS6115074-05mL MyBiosource 0.5(mL 965 EUR

An omics investigation into chronic widespread musculoskeletal pain reveals epiandrosterone sulfate as a potential biomarker.

Chronic widespread musculoskeletal pain (CWP) is common, having a population prevalence of 10%. This study aimed to define the biological basis of the CWP/body mass association by using a systems biology approach. Adult female twins (n = 2444) from the TwinsUK registry who had extensive clinical, anthropometric, and “omic” data were included. Nontargeted metabolomics screening including 324 metabolites was carried out for CWP and body composition using dual-energy X-ray absorptiometry. The biological basis of these associations was explored through a genome-wide association study and replicated in an independent population sample (Cooperative Health Research in the Region of Augsburg [KORA] study, n = 2483). A causal role for the genetic variants identified was sought in CWP using a Mendelian randomisation study design.
Fat mass/height2 was the body composition variable most strongly associated with CWP (TwinsUK: P = 2.4 × 10(-15) and KORA: P = 1.59 × 10(-10)). Of 324 metabolites examined, epiandrosterone sulfate (EAS) was highly associated with both CWP (P = 1.05 × 10(-09) in TwinsUK and P = 3.70 × 10(-06) in KORA) and fat mass/height2. Genome-wide association study of EAS identified imputed single nucleotide polymorphism rs1581492 at 7q22.1 to be strikingly associated with EAS levels (P ≤ 2.49 × 10(-78)), and this result was replicated in KORA (P = 2.12 × 10(-9)). Mendelian randomization by rs1581492 genotype showed that EAS is unlikely to be causally related to CWP. Using an agnostic omics approach to focus on the association of CWP with body mass index, we have confirmed a steroid hormone association and identified a genetic variant upstream of the CYP genes, which likely controls this response. This study suggests that steroid hormone abnormalities result from pain rather than causing it, and EAS may provide a biomarker that identifies subgroups at risk of CWP.

Involvement of c-Jun N-terminal kinase 2 (JNK2) in Endothelin–1 (ET–1) Mediated Neurodegeneration of Retinal Ganglion Cells

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Purpose: The goal of this study was to determine whether JNK2 played a causative role in endothelin-mediated loss of RGCs in mice.
Methods: JNK2-/- and wild type (C57BL/6) mice were intravitreally injected in one eye with 1 nmole of ET-1, whereas the contralateral eye was injected with the vehicle. At two time points (two hours and 24 hours) after the intravitreal injections, mice were euthanized, and phosphorylated c-Jun was assessed in retinal sections. In a separate set of experiments, JNK2-/- and wild type mice were intravitreally injected with either 1 nmole of ET-1 or its vehicle and euthanized seven days after injection. Retinal flat mounts were stained with antibodies to the RGC marker, Brn3a, and surviving RGCs were quantified. Axonal degeneration was assessed in paraphenylenediamine stained optic nerve sections.
Results: Intravitreal ET-1 administration produced a significant increase in immunostaining for phospho c-Jun in wild type mice, which was appreciably lower in the JNK2 -/- mice. A significant (P < 0.05) 26% loss of RGCs was found in wild type mice, seven days after injection with ET-1. JNK2-/- mice showed a significant protection from RGC loss following ET-1 administration, compared to wild type mice injected with ET-1. A significant decrease in axonal counts and an increase in the collapsed axons was found in ET-1 injected wild type mice eyes.
Conclusions: JNK2 appears to play a major role in ET-1 mediated loss of RGCs in mice. Neuroprotective effects in JNK2-/- mice following ET-1 administration occur mainly in the soma and not in the axons of RGCs.

The ring size of monocyclic <em>ET</em>-<em>1</em> controls selectivity and signaling efficiency at both <em>endothelin</em> receptor subtypes

Cardiovascular diseases (CVDs) like hypertension are a major cause for death worldwide. In the cardiovascular tissue, the endothelin system-consisting of the receptor subtypes A (ETA R) and B (ETB R) and the mixed agonist endothelin 1 (ET-1)-is a major key player in the regulation of vascular tone and blood pressure. Tight control of this system is required to maintain homeostasis; otherwise, the endothelin system can cause severe CVDs like pulmonary artery hypertension. The high sequence homology between both receptor subtypes limits the development of novel and selective ligands. Identification of small differences in receptor-ligand interactions and determination of selectivity constraints are crucial to fine-tune ligand properties and subsequent signaling events.
Here, we report on novel ET-1 analogs and their detailed pharmacological characterization. We generated simplified ET-1-derived monocyclic peptides to provide an accessible synthesis route. By detailed in vitro characterization, we demonstrated that both G protein signaling and the subsequent arrestin recruitment of activated ETB R remain intact, whereas activation of the ETA R depends on the intramolecular ring size. Increasing of the intramolecular ring structure reduces activity at the ETA R and shifts the peptide toward ETB R selectivity. All ET-1 analogs displayed efficient ETB R-mediated signaling by G protein activation and arrestin 3 recruitment. Our study provides in-depth characterization of the ET-1/ETA R and ET-1/ETB R interactions, which has the potential for future development of endothelin-based drugs for CVD treatment. By identification of Lys9 for selective labeling, novel analogs for peptide-mediated shuttling by ET-1 are proposed.

<em>Endothelin</em>-<em>1</em> Decreases the Expression of Ephrin-A and B Subtypes in Cultured Rat Astrocytes through <em>ET</em> <sub>B</sub> Receptors

Ephrin family proteins are cell surface molecules that regulate several cellular functions through cell-cell interactions. During nervous tissue repair after injury, the expression of ephrin subtypes in astrocytes is altered, affecting the axonal elongation and migration of neuronal precursors. However, the mechanism regulating the expression of ephrin subtypes in astrocytes has not been investigated. Herein, we studied the effects of endothelin-1 (ET-1) on the expression of ephrin subtypes in cultured rat astrocytes. Our results showed that ET-1 (100 nM) treatment for 1-24 h reduced the expression of ephrin-A2, -A4, -B2, and -B3 mRNA and protein in astrocytes, whereas the expression of ephrin-A1, -A3, -A5, and -B1 mRNA were not affected.
Sarafotoxin S6c, a selective ETB receptor agonist, decreased the expression of ephrin-A2, -A4, -B2, and -B3 in cultured astrocytes. The decrease in ephrin-A2, -A4, -B2, and -B3 expression by ET-1 treatment was reduced in the presence of BQ788, an ETB receptor antagonist, while FR139317, an ETA receptor antagonist, had no effects. These results suggest that ET-1 is a signaling molecule that downregulates ephrin-A2, -A4, -B2, and -B3 expression in astrocytes.

C-type natriuretic peptide (CNP)/Guanylate cyclase B (GC-B) system and <em>endothelin</em>-<em>1</em>(<em>ET</em>-<em>1</em>)/<em>ET</em> receptor A and B system in human vasculature.

In order to assess the physiological and clinical implications of C-type natriuretic peptide (CNP)/guanylyl cyclase B (GC-B) system in the human vasculature, we have examined gene expressions of CNP and its receptor, GC-B, in human vascular endothelial cells (ECs) and smooth muscle cells (SMCs) and have also compared endothelin-1(ET-1)/endothelin receptor-A (ETR-A) and endothelin receptor-B (ETR-B) system in human aortic ECs (HAECs) and vascular SMCs (HSMCs) in vitro. We also examined these gene expressions in human embryonic stem (ES)/induced pluripotent stem cell (iPS)-derived ECs and mural cells (MCs).
A little but significant amount of mRNA encoding CNP was detected in both human ES-derived ECs and HAECs. Substantial amount of GC-B was expressed in both ECs (iPS-derived ECs and HAECs) and SMCs (iPS-derived MCs and HSMCs). ET-1 was expressed solely in ECs. ETR-A was expressed in SMCs, while ETR-B was expressed in ECs. These results indicate the existence of vascular CNP/GC-B system in the human vascular wall, indicating the evidence for clinical implication of CNP/GC-B system in concert with ET-1/ETR-A and ETR-B system in the human vasculature.

Downregulation of endothelial nitric oxide synthase (eNOS) and <em>endothelin</em>-<em>1</em> (<em>ET</em>-<em>1</em>) in a co-culture system with human stimulated X-linked CGD neutrophils.

Phagocytes in patients with chronic granulomatous disease (CGD) do not generate reactive oxidative species (ROS), whereas nitric oxide (NO) production is increased in response to the calcium ionophore A23187 in CGD phagocytes compared with healthy phagocytes. Recently, patients with X-linked CGD (X-CGD) have been reported to show higher flow-mediated dilation, suggesting that endothelial cell function is affected by NO production from phagocytes. We studied NOS3 and EDN1 mRNA and protein expression in human umbilical vein endothelial cells (HUVECs) in a co-culture system with neutrophils from X-CGD patients. HUVECs were co-cultured for 30 minutes with human neutrophils from X-CGD or healthy participants in response to A23187 without cell-to-cell contact. The expression of NOS3 and EDN1 mRNA in HUVECs was quantified by real-time polymerase chain reaction.
Moreover, we demonstrated the protein expression of eNOS, ET-1, and NFκB p65, including phosphorylation at Ser1177 of eNOS and Ser536 of NFκB p65. Neutrophils from X-CGD patients showed significantly higher NO and lower H2O2 production in response to A23187 than healthy neutrophils in vitro. Compared with healthy neutrophils, X-CGD neutrophils under A23187 stimulation exhibited significantly increased NO and decreased H2O2, and promoted downregulated NOS3 and EDN1 expression in HUVECs. The total expression and phosphorylation at Ser1177 of eNOS and ET-1 expression were significantly decreased in HUVECs co-cultures with stimulated X-CGD neutrophils. Also, phosphorylation at Ser536 of NFκB p65 were significantly decreased. In conclusions, eNOS and ET-1 significantly down-regulated in co-culture with stimulated X-CGD neutrophils through their excessive NO and the lack of ROS production. These findings suggest that ROS generated from neutrophils may mediate arterial tone affecting eNOS and ET-1 expression via their NO and ROS production.

DHEA-S production capacity in relation to perceived prolonged stress

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We and other research groups have previously described that levels of the anabolic hormone dehydroepiandrosterone sulfate (DHEA-S) are lowered in individuals who report prolonged stress. We have also shown that the DHEA-S production capacity during acute stress is attenuated in individuals reporting high prolonged stress. This study aimed to further investigate the DHEA and DHEA-S production capacity in relation to prolonged stress. Eighty-one healthy participants in the age 20-50 years old were included in the study and divided into a low stress (n = 45) and a high stress group (n = 36) according their response to a single question regarding perceived stress during the preceding month.
They underwent the Trier Social Stress Test while blood samples were drawn before, during and after the stress test. The concentration of DHEA, DHEA-S, cortisol and ACTH was measured. The results showed that the high stress group exhibited a significantly lower response of DHEA-S (40% lower) than the low stress group, while DHEA, cortisol and ACTH responses did not differ between the groups. Reduced DHEA-S production may constitute one of the links between stress and poor health.

Inter-individual differences in pain anticipation and pain perception in migraine: Neural correlates of migraine frequency and cortisol-to-dehydroepiandrosterone sulfate (DHEA–S) ratio

Previous studies targeting inter-individual differences in pain processing in migraine mainly focused on the perception of pain. Our main aim was to disentangle pain anticipation and perception using a classical fear conditioning task, and investigate how migraine frequency and pre-scan cortisol-to-dehydroepiandrosterone sulfate (DHEA-S) ratio as an index of neurobiological stress response would relate to neural activation in these two phases. Functional Magnetic Resonance Imaging (fMRI) data of 23 participants (18 females; mean age: 27.61± 5.36) with episodic migraine without aura were analysed.
We found that migraine frequency was significantly associated with pain anticipation in brain regions comprising the midcingulate and caudate, whereas pre-scan cortisol-to DHEA-S ratio was related to pain perception in the pre-supplementary motor area (pre-SMA). Both results suggest exaggerated preparatory responses to pain or more general to stressors, which may contribute to the allostatic load caused by stressors and migraine attacks on the brain.

Circulating DHEA–S levels and major cardiovascular outcomes in chronic Chagas cardiomyopathy: A prospective cohort study

Objective: To analyze the association of circulating dehydroepiandrosterone sulfate (DHEA-S) levels with cardiovascular outcomes in patients with chronic Chagas cardiomyopathy (CCM) diagnosis.
Background: DHEA-S is among the main endogenous steroid hormones. Some studies have suggested a relevant role of this hormone in infections and the setting of CCM. Nevertheless, no study has evaluated the prognostic role of DHEA-S in CCM patients.
Methods: Prospective cohort study. Patients with CCM and reduced ejection fraction were included. We explored the association of DHEA-S levels with NT-proBNP levels and echocardiographic variables using linear regression models. Next, by using Cox Proportional Hazard models, we examined whether levels of DHEA-S could predict a composite outcome (CO) including all-cause mortality, cardiac transplantation, and implantation of a left ventricular assist device (LVAD).
Results: Seventy-four patients were included (59% males, median age: 64 years). After adjustment for confounding factors, high DHEA-S levels were associated with better LVEF, lower left atrium volume, end-systolic volume of the left ventricle and lower NT-proBNP levels. 43% of patients experienced the CO during a median follow-up of 40 months. Increased levels of DHEA-S were associated with a lower risk of developing the CO (HR 0.43; 95%CI 0.21-0.86). Finally, adding DHEA-S to the multivariate model did not improve the prediction of the CO, but substituting NT-proBNP in the model with DHEA-S showed similar performance.
Conclusions: In patients with CCM, higher DHEA-S levels were associated with lower mortality, heart transplantation, and LVAD implantation. Further larger studies are required to confirm our results and assess causality.

Comparative Profiling of Salivary Cortisol and Salivary DHEA–S Among Healthy Pregnant and Non-Pregnant Women

  • During pregnancy, circulatory cortisol levels increase, remaining steady over the second-third trimester. In contrast, profile of salivary cortisol during pregnancy is debatable, more influenced by factors like time of sample collection in the day. Circulatory DHEA-S decrease by at least 50% over the second-third trimester of pregnancy. However, profile of salivary DHEA-S is unclear. Objective was to determine changes in salivary cortisol and DHEA-S in healthy pregnant women, compared to non-pregnant women during late morning-early afternoon sampling to avoid fluctuations associated with other times.
  • Pregnant women in their second-third trimester prospectively (n=500) and non-pregnant women (n=133) were enrolled in study with informed consent. Live birth outcome with no pregnancy complications and≥2.5 Kg infant birth weight were included. Concentrations of salivary cortisol and DHEA-S were determined through ELISA assays. Compared to non-pregnant women, pregnant women demonstrated significant increases in salivary cortisol [median (interquartile range)=4.2 (5.1) nmol/l vs. 17.2 (13.9) nmol/l, p<0.001] and salivary DHEA-S median (interquartile range)=2.7 (2.9) nmol/l vs. 3.8 (3.2) nmol/l, p<0.001).
  • Consistently, quartile scores representing higher levels of salivary cortisol and DHEA-S concentrations demonstrated significant association with pregnancy. Quartile scores representing higher salivary cortisol/DHEA-S ratio demonstrated significant association with pregnancy. Study suggests the indicated time range of saliva sampling might best parallel the established profile of circulatory cortisol in pregnant women. However, unlike cortisol, study indicates that the salivary DHEA-S profile is distinct from the well-known profile of circulatory DHEA-S during pregnancy. A combinatorial approach involving both salivary and circulatory compartments could provide comprehensive picture of DHEA-S and hypothalamus-pituitary-adrenal axis during pregnancy.

DHEA-S
DE1562 Demeditec Diagnostics 96 95 EUR

DHEA-S
GWB-DF2891 GenWay Biotech 1x96 Assays Ask for price

DHEA-S
GWB-C990DB GenWay Biotech 1x96 Assays Ask for price

DHEA-S
MBS340681-01mg MyBiosource 0.1mg 1050 EUR

DHEA-S
MBS340681-1mg MyBiosource 1mg 3920 EUR

DHEA-S
MBS340681-5x1mg MyBiosource 5x1mg 17460 EUR

DHEA-S
K00203 DIALAB 96 Well Ask for price

DHEA-S ELISA
GWB-BQK1B7 GenWay Biotech 96 Well Plate Ask for price

DHEA-S ELISA
E22-HC9024 EnoGene 96T 495 EUR

DHEA-S ELISA
MBS8579258-5x96Tests MyBiosource 5x96Tests 2735 EUR

DHEA-S ELISA
MBS8579258-96Tests MyBiosource 96Tests 595 EUR

DHEA-S Antibody
GWB-F09C57 GenWay Biotech 0.1 ml Ask for price

DHEA-S ELISA Kit
MBS495539-5x96Tests MyBiosource 5x96Tests 1555 EUR

DHEA-S ELISA Kit
MBS495539-96Tests MyBiosource 96Tests 340 EUR

DHEA-S ELISA Kit
MBS580010-5x96StripWells MyBiosource 5x96-Strip-Wells 1320 EUR

DHEA-S ELISA Kit
MBS580010-96StripWells MyBiosource 96-Strip-Wells 330 EUR

DHEA-S ELISA Kit
MBS580169-5x96StripWells MyBiosource 5x96-Strip-Wells 1320 EUR

DHEA-S ELISA Kit
MBS580169-96StripWells MyBiosource 96-Strip-Wells 305 EUR

DHEA-S Standard, 350UL
C201-350UL Arbor Assays 350UL 207 EUR

DHEA-S Standard, 70UL
C201-70UL Arbor Assays 70UL 85 EUR

DHEA-S (human) ELISA Kit
K7428-100 Biovision each 940.8 EUR

DHEA-S ELISA KIT|Human
EF006758 Lifescience Market 96 Tests 826.8 EUR

DHEA-S(Dehydroepiandrosteronesulfate)ELISAKit
ELK8370-48T ELK Biotech 48T 206 EUR

DHEA-S(Dehydroepiandrosteronesulfate)ELISAKit
ELK8370-96T ELK Biotech 96T 294 EUR

RealScreen DHEA-S ELISA Kit
EL102-096 GenDepot 96T 1161.6 EUR

DHEA-S ELISA Kit (1 Plate)
K054-H1 Arbor Assays 1x96 well plate 444 EUR

DHEA-S ELISA Kit (5 Plate)
K054-H5 Arbor Assays 5x96 well plate 1775 EUR

Anti-Dehydroepiandrosterone Sulphate (DHEA-S)
MBS340106-10mg MyBiosource 10mg 1920 EUR

Anti-Dehydroepiandrosterone Sulphate (DHEA-S)
MBS340106-1mg MyBiosource 1mg 615 EUR

Anti-Dehydroepiandrosterone Sulphate (DHEA-S)
MBS340106-5x10mg MyBiosource 5x10mg 8470 EUR

Comparative profiling of prenatal cortisol and DHEA–S among pregnant women with poor birth outcome and pregnant women with normal birth outcome

Context: Cortisol and dehydroepiandrosterone-sulfate (DHEA-S) are indispensable hormones for normal pregnancy. It is unclear if these hormones, specifically DHEA-S can offer value for predicting poor birth outcome.
Objective: To compare prenatal cortisol and DHEA-S levels among pregnant women with normal or poor birth outcome.
Design-patients: Plasma and saliva were collected prospectively from women in second-third trimester of pregnancy. Women with normal birth outcome (NBO), (n=501) included live birth, no pregnancy complications and ≥ 2.5Kg infant birth weight. Women with poor birth outcome included adverse birth (ABO), (n=50) or low birth weight outcome (LBW), (n=147).
Measurements: ELISA was performed to measure hormone concentrations in plasma and saliva.
Results: Circulatory-DHEA-S levels in pregnant women with ABO were higher than women with NBO (p=0.043). Among ABO, only stillbirth cases demonstrated significant increase in circulatory-DHEA-S levels (p=0.006). Circulatory and salivary cortisol/DHEA-S ratio was lower among women with stillbirth (p=0.004) and ABO outcome (p=0.043) respectively compared to women with NBO. Consistently, increased odds of ABO were observed in pregnant women with highest circulatory-DHEA-S levels (odds ratio quartile score 1 vs 4, 2.79, p=0.027) and lowest salivary cortisol/DHEA-S ratio (score 4 vs 2, 2.83, p=0.025). Increased odds of stillbirth outcome were observed in pregnant women with highest circulatory-DHEA-S levels (odds ratio quartile score 1 vs 4, 8.47, p=0.046) and lowest circulatory cortisol/DHEA-S ratio (score 4 vs 1, 4.803, p=0.048). Associations remained significant after adjusting for confounders. Women with LBW did not demonstrate significant changes in cortisol or DHEA-S levels.
Conclusion: Prenatal measurement of DHEA-S or cortisol/DHEA-S ratio may offer significant value for predicting adverse birth, specifically stillbirth outcome.

Nitrate Is an Environmental Cue in the Gut for Salmonella enterica Serovar Typhimurium Biofilm Dispersal through Curli Repression and Flagellum Activation via Cyclic-di-GMP Signaling

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Curli, a major component of the bacterial biofilms in the intestinal tract, activates pattern recognition receptors and triggers joint inflammation after infection with Salmonella enterica serovar Typhimurium. The factors that allow S. Typhimurium to disperse from biofilms and invade the epithelium to establish a successful infection during acute inflammation remain unknown. Here, we studied S. Typhimurium biofilms in vitro and in vivo to understand how the inflammatory environment regulates the switch between multicellular and motile S. Typhimurium in the gut. We discovered that nitrate generated by the host is an environmental cue that induces S. Typhimurium to disperse from the biofilm. Nitrate represses production of an important biofilm component, curli, and activates flagella via the modulation of intracellular cyclic-di-GMP levels. We conclude that nitrate plays a central role in pathogen fitness by regulating the sessile-to-motile lifestyle switch during infection.
IMPORTANCE Recent studies provided important insight into our understanding of the role of c-di-GMP signaling and the regulation of enteric biofilms. Despite an improved understanding of how c-di-GMP signaling regulates S. Typhimurium biofilms, the processes that affect the intracellular c-di-GMP levels and the formation of multicellular communities in vivo during infections remain unknown. Here, we show that nitrate generated in the intestinal lumen during infection with S. Typhimurium is an important regulator of biofilm formation in vivo.

Second Messenger 2’3′-cyclic GMP-AMP (2’3′-cGAMP):Synthesis, transmission, and degradation

Cyclic GMP-AMP synthase (cGAS) senses foreign DNA to produce 2’3′-cyclic GMP-AMP (2’3′-cGAMP). 2’3′-cGAMP is a second messenger that binds and activates the adaptor protein STING, which triggers the innate immune response. As a STING agonist, the small molecule 2’3′-cGAMP plays pivotal roles in antiviral defense and has adjuvant applications, and anti-tumor effects. 2’3′-cGAMP and its analogs are thus putative targets for immunotherapy and are currently being testedin clinical trials to treat solid tumors.
However, several barriers to further development have emerged from these studies, such as evidence of immune and inflammatory side-effects, poor pharmacokinetics, and undesirable biodistribution. Here, we review the status of 2’3′-cGAMP research and outline the role of 2’3′-cGAMP in immune signaling, adjuvant applications, and cancer immunotherapy, as well as various 2’3′-cGAMP detection methods.

A pGpG-specific phosphodiesterase regulates cyclic di-GMP signaling in Vibrio cholerae

The bacterial second messenger cyclic diguanylate monophosphate (c-di-GMP) controls various cellular processes, including motility, toxin production and biofilm formation. c-di-GMP is enzymatically synthesized by GGDEF domain-containing diguanylate cyclases and degraded by HD-GYP domain-containing phosphodiesterases (PDEs) to 2 GMP or by EAL domain-containing PDE-As to 5′-phosphoguanylyl-(3′,5′)-guanosine (pGpG). Since excess pGpG feedback inhibits PDE-A activity and thereby can lead to the uncontrolled accumulation of c-di-GMP, a PDE that degrades pGpG to 2 GMP (PDE-B) has been presumed to exist.
To date, the only enzyme known to hydrolyze pGpG is Oligoribonuclease Orn, which degrades all kinds of oligoribonucleotides. Here, we identified a pGpG-specific PDE, which we named PggH, using biochemical approaches in the gram-negative bacteria Vibrio cholerae. Biochemical experiments revealed that PggH exhibited specific PDE activity only toward pGpG, thus differing from the previously reported Orn. Furthermore, the high-resolution structure of PggH revealed the basis for its PDE activity and narrow substrate specificity. Finally, we propose that PggH could modulate the activities of PDE-As and the intracellular concentration of c-di-GMP, resulting in phenotypic changes including in biofilm formation.

The Campylobacter jejuni Response Regulator and Cyclic-Di-GMP Binding CbrR Is a Novel Regulator of Flagellar Motility

A leading cause of bacterial gastroenteritis, Campylobacter jejuni is also associated with broad sequelae, including extragastrointestinal conditions such as reactive arthritis and Guillain-Barré Syndrome (GBS). CbrR is a C. jejuni response regulator that is annotated as a diguanylate cyclase (DGC), an enzyme that catalyzes the synthesis of c-di-GMP, a universal bacterial second messenger, from GTP. In C. jejuni DRH212, we constructed an unmarked deletion mutant, cbrR–, and complemented mutant, cbrR+. Motility assays indicated a hyper-motile phenotype associated with cbrR–, whereas motility was deficient in cbrR+.
The overexpression of CbrR in cbrR+ was accompanied by a reduction in expression of FlaA, the major flagellin. Biofilm assays and scanning electron microscopy demonstrated similarities between DRH212 and cbrR–; however, cbrR+ was unable to form significant biofilms. Transmission electron microscopy showed similar cell morphology between the three strains; however, cbrR+ cells lacked flagella. Differential radial capillary action of ligand assays (DRaCALA) showed that CbrR binds GTP and c-di-GMP. Liquid chromatography tandem mass spectrometry detected low levels of c-di-GMP in C. jejuni and in E. coli expressing CbrR. CbrR is therefore a negative regulator of FlaA expression and motility, a critical virulence factor in C. jejuni pathogenesis.

The Regulatory Network of Cyclic GMP-AMP Synthase-Stimulator of Interferon Genes Pathway in Viral Evasion

Virus infection has been consistently threatening public health. The cyclic GMP-AMP synthase (cGAS)-Stimulator of Interferon Genes (STING) pathway is a critical defender to sense various pathogens and trigger innate immunity of mammalian cells. cGAS recognizes the pathogenic DNA in the cytosol and then synthesizes 2’3′-cyclic GMP-AMP (2’3’cGAMP). As the second messenger, cGAMP activates STING and induces the following cascade to produce type I interferon (IFN-I) to protect against infections. However, viruses have evolved numerous strategies to hinder the cGAS-STING signal transduction, promoting their immune evasion.
Here we outline the current status of the viral evasion mechanism underlying the regulation of the cGAS-STING pathway, focusing on how post-transcriptional modifications, viral proteins, and non-coding RNAs involve innate immunity during viral infection, attempting to inspire new targets discovery and uncover potential clinical antiviral treatments.

cGMP (cyclic GMP)
MBS6507358-005mL MyBiosource 0.05mL 835 EUR

cGMP (cyclic GMP)
MBS6507358-5x005mL MyBiosource 5x0.05mL 3600 EUR

Cyclic GMP (TBAOH)
HY-113469B MedChemExpress 10 mg 70.35 EUR

Cyclic GMP (sodium)
HY-113469A MedChemExpress 10 mg 70.35 EUR

Cyclic GMP Antibody
abx022640-02ml Abbexa 0.2 ml 1128 EUR

Cyclic GMP Antibody
GWB-DFA25F GenWay Biotech 2 ml Ask for price

Cyclic GMP Antibody
GWB-BA5CC2 GenWay Biotech 0.2 ml Ask for price

Cyclic GMP EIA Kit
SKT-211-96 Stressmarq 1 plate of 96 wells 568.8 EUR

Sheep anti Cyclic GMP
MBS316214-02mL MyBiosource 0.2mL 1235 EUR

Sheep anti Cyclic GMP
MBS316214-5x02mL MyBiosource 5x0.2mL 5380 EUR

SHEEP ANTI CYCLIC GMP
MBS222513-2mL MyBiosource 2mL 455 EUR

SHEEP ANTI CYCLIC GMP
MBS222513-5x2mL MyBiosource 5x2mL 1870 EUR

Cyclic GMP (Direct) ELISA
MBS494508-5x96Tests MyBiosource 5x96Tests 3410 EUR

Cyclic GMP (Direct) ELISA
MBS494508-96Tests MyBiosource 96Tests 740 EUR

Cyclic GMP Standard, 125UL
C080-125UL Arbor Assays 125UL 85 EUR

Cyclic GMP Standard, 350UL
C080-350UL Arbor Assays 350UL 207 EUR

Cyclic GMP Standard, 625UL
C080-625UL Arbor Assays 625UL 207 EUR

Cyclic GMP Standard, 70UL
C080-70UL Arbor Assays 70UL 85 EUR

Anti- Cyclic GMP Antibody
GWB-FDE020 GenWay Biotech 1000 TESTS Ask for price

Cyclic GMP Polyclonal Antibody
MBS194212-02mL MyBiosource 0.2mL 560 EUR

Cyclic GMP Polyclonal Antibody
MBS194212-5x02mL MyBiosource 5x0.2mL 2510 EUR

cGMP(Cyclic GMP)ELISA Kit
MBS2516175-10x96Tests MyBiosource 10x96Tests 3265 EUR

cGMP(Cyclic GMP)ELISA Kit
MBS2516175-24Tests MyBiosource 24Tests 225 EUR

cGMP(Cyclic GMP)ELISA Kit
MBS2516175-48Test MyBiosource 48Test 350 EUR

cGMP(Cyclic GMP)ELISA Kit
MBS2516175-5x96Test MyBiosource 5x96Test 1655 EUR

cGMP(Cyclic GMP)ELISA Kit
MBS2516175-96Tests MyBiosource 96Tests 395 EUR

cGMP(Cyclic GMP) ELISA Kit
YPJ1147-48wellsplate UpingBio 48 wells plate 180 EUR

cGMP(Cyclic GMP) ELISA Kit
YPJ1147-96wellsplate UpingBio 96 wells plate 220 EUR

BldD-based bimolecular fluorescence complementation for in vivo detection of the second messenger cyclic di-GMP

The widespread bacterial second messenger bis-(3′-5′)-cyclic diguanosine monophosphate (c-di-GMP) is an important regulator of biofilm formation, virulence and cell differentiation. C-di-GMP-specific biosensors that allow detection and visualization of c-di-GMP levels in living cells are key to our understanding of how c-di-GMP fluctuations drive cellular responses. Here, we describe a novel c-di-GMP biosensor, CensYBL, that is based on c-di-GMP-induced dimerization of the effector protein BldD from Streptomyces resulting in bimolecular fluorescence complementation of split-YPet fusion proteins.
As a proof-of-principle, we demonstrate that CensYBL is functional in detecting fluctuations in intracellular c-di-GMP levels in the Gram-negative model bacteria Escherichia coli and Salmonella enterica serovar Typhimurium. Using deletion mutants of c-di-GMP diguanylate cyclases and phosphodiesterases, we show that c-di-GMP dependent dimerization of CBldD-YPet results in fluorescence complementation reflecting intracellular c-di-GMP levels. Overall, we demonstrate that the CensYBL biosensor is a user-friendly and versatile tool that allows to investigate c-di-GMP variations using single-cell and population-wide experimental set-ups.

Urinary biomarkers for the detection of ovarian cancer: A systematic review

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Currently, the only definitive method for diagnosing ovarian cancer involves histological examination of tissue obtained at time of surgery or by invasive biopsy. Blood has traditionally been the biofluid of choice in ovarian cancer biomarker discovery; however, there has been a growing interest in exploring urinary biomarkers, particularly as it is non-invasive. In this systematic review, we present the diagnostic accuracy of urinary biomarker candidates for the detection of ovarian cancer. A comprehensive literature search was performed using the MEDLINE/PubMed and EMBASE, up to 1 st April 2021. All included studies reported the diagnostic accuracy using sensitivity and/or specificity and/or receiver operating characteristics (ROC) curve. Risk of bias and applicability of included studies were assessed using the QUADAS-2 tool.
Twenty seven studies were included in the narrative synthesis. Protein/peptide biomarkers were most commonly described (n=18), with seven studies reporting composite scores of multiple protein-based targets. The most frequently described urinary protein biomarker was HE4 (n=5), with three studies reporting a sensitivity and specificity >80%. Epigenetic (n=1) and metabolomic/organic compound biomarkers (n=8) were less commonly described. Overall, six studies achieved a sensitivity and specificity of >90% and/or an AUC >0.9. Evaluation of urinary biomarkers for the detection of ovarian cancer is a dynamic and growing field. Currently, the most promising biomarkers are those that interrogate metabolomic pathways and organic compounds, or quantify multiple proteins. Such biomarkers require external validation in large, prospective observational studies before they can be implemented into clinical practice.

Rapid electrostatic DNA enrichment for sensitive detection of Trichomonas vaginalis in clinical urinary samples

Estimated to be the most common non-viral sexually transmitted infection globally, Trichomonas vaginalis (TV) can lead to pelvic inflammatory disease, pregnancy complications, and increased risk of acquiring and transmitting HIV. Once diagnosed, TV infection can be treated with oral antibiotics; however, infected individuals are often asymptomatic and do not seek treatment. The WHO and others have identified a need for point-of-care tests to expand access to TV testing and screening; ideal test characteristics include high sensitivity and specificity and the ability to use urine as a sample type, rather than invasively collected swab samples. Here, we report on a proof-of-concept prototype for rapid, electrostatic enrichment of DNA from urine samples and demonstrate the use of large volumes of urine to increase sensitivity of downstream nucleic acid amplification testing.
We developed an internally controlled thermophilic helicase-dependent amplification (tHDA) assay with lateral flow immunoassay readout and demonstrate that this tHDA assay can be performed directly on our DNA capture filter. We validated our method using clinical urine samples with qPCR-quantified TV loads. Using 62 clinical urine samples and a simple sample processing device, our tHDA assay displayed 96.6% sensitivity and 100% specificity. Our analytical limit of detection was found to be approximately 7 genomic equivalents of TV DNA per mL of sample when 1 mL of sample was tested, comparable to existing isothermal tests for TV. Using large-volume simulated samples (40 mL of buffered urine with spiked-in TV DNA), we also demonstrated that sensitivity could be improved 28-fold to 0.25 genomic equivalents of TV DNA per mL, with a sample processing time of only 2 minutes.

Nanomaterials-Based Urinary Extracellular Vesicles Isolation and Detection for Non-invasive Auxiliary Diagnosis of Prostate Cancer

Extracellular vesicles (EVs) are natural nanoparticles secreted by cells in the body and released into the extracellular environment. They are associated with various physiological or pathological processes, and considered as carriers in intercellular information transmission, so that EVs can be used as an important marker of liquid biopsy for disease diagnosis and prognosis. EVs are widely present in various body fluids, among which, urine is easy to obtain in large amount through non-invasive methods and has a small dynamic range of proteins, so it is a good object for studying EVs. However, most of the current isolation and detection of EVs still use traditional methods, which are of low purity, time consuming, and poor efficiency; therefore, more efficient and highly selective techniques are urgently needed. Recently, inspired by the nanoscale of EVs, platforms based on nanomaterials have been innovatively explored for isolation and detection of EVs from body fluids.
These newly developed nanotechnologies, with higher selectivity and sensitivity, greatly improve the precision of isolation target EVs from urine. This review focuses on the nanomaterials used in isolation and detection of urinary EVs, discusses the advantages and disadvantages between traditional methods and nanomaterials-based platforms, and presents urinary EV-derived biomarkers for prostate cancer (PCa) diagnosis. We aim to provide a reference for researchers who want to carry out studies about nanomaterial-based platforms to identify urinary EVs, and we hope to summarize the biomarkers in downstream analysis of urinary EVs for auxiliary diagnosis of PCa disease in detail.

Rational Design of Phe-BODIPY Amino Acids as Fluorogenic Building Blocks for Peptide-based Detection of Urinary Tract Candida Infections

Fungal infections caused by Candida species are among the most prevalent in hospitalized patients. However, current methods for the detection of Candida fungal cells in clinical samples rely on time-consuming assays that hamper rapid and reliable diagnosis. Herein, we describe the rational development of new Phe-BODIPY amino acids as small fluorogenic building blocks and their application to generate fluorescent antimicrobial peptides for rapid labelling of Candida cells in urine.
We have used computational methods to analyse the fluorogenic behaviour of BODIPY-substituted aromatic amino acids and performed bioactivity and confocal microscopy experiments in different strains to confirm the utility and versatility of peptides incorporating Phe-BODIPYs. Finally, we have designed a simple and sensitive fluorescence-based assay for the detection of Candida albicans in human urine samples.

Creatinine Urinary Detection Kit (2 Plate)
K002-H1 Arbor Assays 2x96 well plates 296 EUR

Creatinine Urinary Detection Kit (10 Plate)
K002-H5 Arbor Assays 10x96 well plates 1182 EUR

OKAU00002-2PLATE - Creatinine Urinary Detection Kit
OKAU00002-2PLATE Aviva Systems Biology 2plate 259 EUR

OKAU00002-10PLATE - Creatinine Urinary Detection Kit
OKAU00002-10PLATE Aviva Systems Biology 10plate 879 EUR

Urine Creatinine Detection Kit
MBS807896-10x96Wells MyBiosource 10x96Wells 1710 EUR

Urine Creatinine Detection Kit
MBS807896-2x96Wells MyBiosource 2x96Wells 375 EUR

Urine Creatinine Detection Kit
SKT-200-192 Eagle Biosciences 1 Kit Ask for price

Exosome Purification and Detection Kit (Urine)
abx290027-25tests Abbexa 25 tests 777.6 EUR

Multi-Species Creatinine Detection Kit for Urine
IMLCRKTBF Innovative research each 387 EUR

Multi-Species Creatinine Detection Kit for Urine
MBS8420179-1Kit MyBiosource 1Kit 555 EUR

Multi-Species Creatinine Detection Kit for Urine
MBS8420179-5x1Kit MyBiosource 5x1Kit 2510 EUR

SEB Detection Kit
6030 Chondrex 1 kit 377 EUR

LPS Detection Kit
6039 Chondrex 1 kit 404 EUR

NGAL (Detection Ab)
abx019242-100ug Abbexa 100 ug 777.6 EUR

DAB Detection Kit
E-IR-R101-1mL Elabscience Biotech 1mL 35 EUR

DAB Detection Kit
E-IR-R101-3mL Elabscience Biotech 3mL 50 EUR

DAB Detection Kit
E-IR-R101-6mL Elabscience Biotech 6mL 85 EUR

DAB Detection Kit
E-IR-R101-each Elabscience Biotech each Ask for price

TLR Detection Set
MBS154739-1Set MyBiosource 1Set 1410 EUR

TLR Detection Set
MBS154739-5x1Set MyBiosource 5x1Set 6515 EUR

NGAL (Detection Ab)
MBS355008-1mg MyBiosource 1mg 570 EUR

NGAL (Detection Ab)
MBS355008-5x1mg MyBiosource 5x1mg 2265 EUR

TLR Detection Set
PSI-1806 ProSci 1 Set 1627.8 EUR

PD1 Detection Set
SD8600 ProSci 1 Set 537.9 EUR

HROS Detection Kit
FLAPF100-2 Cell Technology 150 Tests 280 EUR

IRAK Detection Set
MBS154722-1Set MyBiosource 1Set 770 EUR

A Prospective Multicenter Trial to Evaluate Urinary Metabolomics for Non-invasive Detection of Renal Allograft Rejection (PARASOL): Study Protocol and Patient Recruitment

Background: In an earlier monocentric study, we have developed a novel non-invasive test system for the prediction of renal allograft rejection, based on the detection of a specific urine metabolite constellation. To further validate our results in a large real-world patient cohort, we designed a multicentric observational prospective study (PARASOL) including six independent European transplant centers. This article describes the study protocol and characteristics of recruited better patients as subjects.
Methods: Within the PARASOL study, urine samples were taken from renal transplant recipients when kidney biopsies were performed. According to the Banff classification, urine samples were assigned to a case group (renal allograft rejection), a control group (normal renal histology), or an additional group (kidney damage other than rejection).
Results: Between June 2017 and March 2020, 972 transplant recipients were included in the trial (1,230 urine samples and matched biopsies, respectively). Overall, 237 samples (19.3%) were assigned to the case group, 541 (44.0%) to the control group, and 452 (36.7%) samples to the additional group. About 65.9% were obtained from male patients, the mean age of transplant recipients participating in the study was 53.7 ± 13.8 years. The most frequently used immunosuppressive drugs were tacrolimus (92.8%), mycophenolate mofetil (88.0%), and steroids (79.3%). Antihypertensives and antidiabetics were used in 88.0 and 27.4% of the patients, respectively. Approximately 20.9% of patients showed the presence of circulating donor-specific anti-HLA IgG antibodies at time of biopsy. Most of the samples (51.1%) were collected within the first 6 months after transplantation, 48.0% were protocol biopsies, followed by event-driven (43.6%), and follow-up biopsies (8.5%). Over time the proportion of biopsies classified into the categories Banff 4 (T-cell-mediated rejection [TCMR]) and Banff 1 (normal tissue) decreased whereas Banff 2 (antibody-mediated rejection [ABMR]) and Banff 5I (mild interstitial fibrosis and tubular atrophy) increased to 84.2 and 74.5%, respectively, after 4 years post transplantation. Patients with rejection showed worse kidney function than patients without rejection.
Conclusion: The clinical characteristics of subjects recruited indicate a patient cohort typical for routine renal transplantation all over Europe. A typical shift from T-cellular early rejections episodes to later antibody mediated allograft damage over time after renal transplantation further strengthens the usefulness of our cohort for the evaluation of novel biomarkers for allograft damage.

Serum creatinine to cystatin C ratio reflects preoperative and early postoperative walking ability in older patients with hip fracture

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Background: The sarcopenia index (SI), calculated as the ratio of serum creatinine to cystatin C levels, reflects skeletal muscle mass and strength. Patients with hip fracture (HF) and sarcopenia have poor functional outcomes, and many require long-term care after surgery. We hypothesized that the SI can predict preoperative and early postoperative functional outcomes.
Methods: Preoperative serum creatinine and cystatin C were measured to calculate the SI for patients with surgically treated HF (n = 130, mean age: 87.8 ± 6.9 years). Walking ability before and 2 weeks after surgery was assessed, and patients were dichotomized into independent and assistance groups. To assess the validity of the SI, we examined its correlation with the quality [computed tomography (CT) value] and quantity (cross-sectional area) of the muscles around the hip on the non-operated side, which were preoperatively measured using CT. Receiver operating characteristic (ROC) analysis was performed to evaluate the prognostic value of the SI.
Results: The SI of the preoperative independent (n = 77) and assistance groups (n = 53) significantly differed (70.2 ± 12.4 and 60.1 ± 9.8, respectively, P < 0.000001). At 2 weeks after surgery, the SI was significantly higher in the independent group (n = 31, 73.0 ± 14.9) than in the assistance group (n = 99, 64.0 ± 10.7, P = 0.0003). In the preoperative independent group, 28 could walk independently after surgery (SI: 74.8 ± 14.0) while 49 required assistance (SI: 67.7 ± 10.6, P = 0.01). For patients with femoral neck fracture (FNF), the SIs were significantly higher in the postoperative independent group (78.6 ± 15.7) than in the postoperative assistance group (63.2 ± 10.9, P = 0.002). Logistic regression analysis showed that the odds ratio (95% confidence interval) of the SI for postoperative walking ability was 0.95 (0.91-0.99, P = 0.03). The correlations of SIs with CT values and cross-sectional areas were as follows: iliopsoas at the apex of the femoral head, r = 0.40, P < 0.001 and r = 0.49, P < 0.001, respectively; rectus femoris at the level of the lessor trochanter, r = 0.26, P = 0.007 and r = 0.37, P < 0.001, respectively. ROC analysis for predicting postoperative walking ability in preoperative independent patients with HF and FNF revealed areas under the curve (95% confidence interval) of 0.63 (0.50-0.76) and 0.80 (0.65-0.96), respectively.
Conclusions: In patients with HF, the SI correlated with preoperative walking ability and could predict postoperative walking ability. Among patients who could walk independently before surgery, those with high SIs could walk independently early in the postoperative period. The SI is beneficial for estimating walking ability in patients with HF.

Relationship Between Serum Uric Acid-to-Creatinine Ratio and the Risk of Metabolic-Associated Fatty Liver Disease in Patients with Type 2 Diabetes Mellitus

Purpose: To investigate the association between serum uric acid-to-creatinine ratio (SUA/Cr) and the risk of developing metabolic-associated fatty liver disease (MAFLD) in patients with type 2 diabetes mellitus (T2DM).
Patients and methods: Overall, 1434 patients with T2DM who were admitted to Hebei General Hospital from January 2019 to December 2019 were selected as the study subjects. According to abdominal ultrasound findings, patients were divided into two groups: MAFLD group and non-MAFLD group. A total of 734 patients were diagnosed with MAFLD. Participants were divided into three study groups according to the SUA/Cr ratio. Chi-square test and one-way analysis of variance were used to perform a comparison between groups. The relationship between SUA/Cr ratio and MAFLD risk was analyzed using correlation analysis and regression analysis. Furthermore, subgroup analyses were performed to verify the robustness of the results.
Results: The detection rate of MAFLD in patients with T2DM was 51.2%, and the detection rate of progressive liver fibrosis in T2DM patients with MAFLD was 36.6%. A significantly higher SUA/Cr ratio was seen in the MAFLD group than in the non-MAFLD group. After adjusting for confounding factors, multivariate logistic regression analysis revealed that the SUA/Cr ratio was an independent risk factor for MAFLD development. Stronger correlations were found in participants with a body mass index ranging between 23 and 28 kg/m2, HbA1C >7%, or female sex.
Conclusion: An elevated SUA/Cr index is independently correlated with an increased risk of MAFLD in Chinese adults with T2DM.

Early prediction of COVID-19-associated Acute Kidney Injury: Are serum NGAL and serum Cystatin C levels better than serum creatinine?

Background: Coronavirus disease-2019 (COVID-19) is associated with a high risk of acute kidney injury (AKI), often requiring renal replacement therapy (RRT). Serum Cystatin C (sCysC) and serum Neutrophil Gelatinase-Associated Lipocalin (sNGAL) are emerging biomarkers for kidney injury, and were suggested to be superior to serum creatinine (sCr) in several clinical settings. Moreover, elevated sCysC is associated with disease severity and mortality in COVID-19. We aimed to assess the utility of sCysC and sNGAL for predicting COVID-19-associated AKI, need for RRT, and need for intensive care unit (ICU) admission, when measured at presentation to the emergency department (ED).
Methods: Patients presenting to the ED with laboratory-confirmed COVID-19 were included. The primary outcome was development of COVID-19-associated AKI, while the secondary outcomes were need for RRT and ICU admission.
Results: Among 52 COVID-19 patients, 22 (42.3%) developed AKI with 8/22 (36.4%) requiring RRT. Both sCr and sCysC demonstrated excellent performance for predicting AKI (AUC, 0.86 and 0.87, respectively) and need for RRT (AUC, 0.94 and 0.95, respectively). sNGAL displayed acceptable performance for predicting AKI (AUC, 0.81) and need for RRT (AUC, 0.87).
Conclusions: SCr and sCysC measured at ED presentation are both highly accurate predictors of AKI and need for RRT, whereas sNGAL demonstrated adequate diagnostic performance. While sCyC was previously shown to be superior to sCr as a diagnostic biomarker of kidney injury in certain etiologies, our findings demonstrate that sCr is comparable to sCyC in the context of predicting COVID-19-associated AKI. Given the high sensitivity of these biomarkers for predicting the need for RRT, and as sCysC is associated with mortality in COVID-19 patients, we recommend their measurement for enabling risk stratification and early intervention.

Creatinine Serum Samples
MBS173604-1Sample MyBiosource 1Sample 310 EUR

Creatinine Serum Samples
MBS173604-5Samples MyBiosource 5Samples 1005 EUR

Creatinine Serum Samples
MBS173604-5x5Samples MyBiosource 5x5Samples 4310 EUR

Creatinine Serum Detection Kit
SKT-217-192 Eagle Biosciences 1 Kit Ask for price

Creatinine Serum Kit (2 Plate)
KB02-H1 Arbor Assays 2x96 well plates 302 EUR

Creatinine Serum Kit (4 Plate)
KB02-H2 Arbor Assays 4x96 well plates 484 EUR

OKAU00065-1PLATE - Creatinine Serum Kit
OKAU00065-1PLATE Aviva Systems Biology 1plate 379 EUR

OKAU00065-2PLATE - Creatinine Serum Kit
OKAU00065-2PLATE Aviva Systems Biology 2plate 269 EUR

OKAU00065-4PLATE - Creatinine Serum Kit
OKAU00065-4PLATE Aviva Systems Biology 4plate 439 EUR

Creatinine Serum Low Sample Volume Kit (384-well Plate)
KB02-H1D Arbor Assays 1x384 well plate 431 EUR

Multi-Species Creatinine Detection Kit for Plasma and Serum
IMLCRKTPS Innovative research each 395 EUR

Multi-Species Creatinine Detection Kit for Plasma and Serum
MBS8420180-1Kit MyBiosource 1Kit 565 EUR

Multi-Species Creatinine Detection Kit for Plasma and Serum
MBS8420180-5x1Kit MyBiosource 5x1Kit 2555 EUR

Serum Creatinine ELISA kit (colorimetric, all species), 96 tests, quantitative
100-300-SCR Alpha Diagnostics 1 kit 343.2 EUR

Serum Creatinine ELISA kit (colorimetric, all species), 2x96 tests, quantitative
100-305-SCR Alpha Diagnostics 1 kit 562.8 EUR

Creatinine
591968 MedKoo Biosciences 25.0g 220 EUR

Creatinine
09626-34 NACALAI TESQUE 5G 11.55 EUR

Creatinine
09626-92 NACALAI TESQUE 25G 25.2 EUR

Creatinine
C781500 Toronto Research Chemicals 10g 74 EUR

Creatinine
CB0328 Bio Basic 5g 68.35 EUR

Creatinine
C271-100MG TOKU-E 100 mg 228.03 EUR

Creatinine
C271-25MG TOKU-E 25 mg 65.03 EUR

The effect of gender-affirming hormone treatment on serum creatinine in transgender and gender-diverse youth: implications for estimating GFR

Background: Equations for estimated glomerular filtration rate (eGFR) based on serum creatinine include terms for sex/gender. For transgender and gender-diverse (TGD) youth, gender-affirming hormone (GAH) treatment may affect serum creatinine and in turn eGFR.
Methods: TGD youth were recruited for this prospective, longitudinal, observational study prior to starting GAH treatment. Data collected as part of routine clinical care were abstracted from the medical record.
Results: For participants designated male at birth (DMAB, N = 92), serum creatinine decreased within 6 months of estradiol treatment (mean ± SD 0.83 ± 0.12 mg/dL to 0.76 ± 0.12 mg/dL, p < 0.001); for participants designated female at birth (DFAB, n = 194), serum creatinine increased within 6 months of testosterone treatment (0.68 ± 0.10 mg/dL to 0.79 ± 0.11 mg/dL, p < 0.001). Participants DFAB treated with testosterone had serum creatinine similar to that of participants DMAB at baseline, whereas even after estradiol treatment, serum creatinine in participants DMAB remained higher than that of participants DFAB at baseline. Compared to reference groups drawn from the National Health and Nutritional Examination Survey, serum creatinine after 12 months of GAH was more similar when compared by gender identity than by designated sex.
Conclusion: GAH treatment leads to changes in serum creatinine within 6 months of treatment. Clinicians should consider a patient’s hormonal exposure when estimating kidney function via eGFR and use other methods to estimate GFR if eGFR based on serum creatinine is concerning.

Cepheid GeneXpert Controls

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Microbiologics, the leading provider of control strains, offers QC panels designed specifically for Cepheid GeneXpert
systems. Our QC Panels feature individually packaged, pre-mixed pools of inactivated control strains recommended by
the instrument manufacturer. The organisms are dried onto a swab to mimic the processing of a patient sample.

Accurate, Reliable Results

  • Recommended for routine QC
  • Protects your reputation by providing documentation and assurance that the laboratory can consistently produce accurate results
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Easy-to-Use

  • No mixing required
  • Individually packaged and color-coordinated labeling to simplify use and avoid cross-contamination
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  • For in vitro diagnostic use (IVD)