Regioselective synthesis, physicochemical properties and anticancer activity of 2-aminomethylated estrone derivatives

The unique estrogen receptor (ER)-independent antiproliferative and apoptotic activity of 2-methoxyestradiol (2ME2) is well known, however, its use has been limited because of its poor oral bioavailability. In this study, novel 2-aminomethylated estrone (E) and estradiol (E2) derivatives structurally related to 2ME2 were synthesized, and their physicochemical properties as well as their in vitro cytotoxic effects were investigated in the hope to find more selective antiproliferative agents with improved pharmacokinetic profile. The target compounds were synthesized from 2-dimethylaminomethylated E obtained regioselectively by a three-component Mannich reaction. Quaternization with methyl iodide followed by reacting the ammonium salt with various dialkyl and alicyclic secondary amines afforded the desired products in good yields.
The reactions proceeded via a 1,4-nucleophilic addition of the applied secondary amines to the ortho-quinone methide (o-QM) intermediates, generated in situ from the salt by base-promoted β-elimination. The compound library has been enlarged with structurally similar E2 analogues obtained by stereoselective reduction and with some 17β-benzylamino derivatives prepared by reductive amination. The potential values of the novel E and E2 derivatives were characterised by means of three different approaches. At the first step compounds were virtually screened using physicochemical parameters. Physicochemical characterization was completed by kinetic solubility and in vitro intestinal-specific permeability measurement. Antiproliferative effects were additionally determined on a panel of malignant and non-cancerous cell lines. The evaluation of the pharmacological profile of the novel E and E2 derivatives was completed with the calculation of lipophilic efficacy (LiPE).

The Role of Estriol and Estrone in Keratoconic Stromal Sex Hormone Receptors

Keratoconus (KC) is a progressive corneal thinning disease that manifests in puberty and worsens during pregnancy. KC onset and progression are attributed to diverse factors that include: environmental, genetics, and hormonal imbalances; however, the pathobiology remains elusive. This study aims to determine the role of corneal stroma sex hormone receptors in KC and their interplay with estrone (E1) and estriol (E3) using our established 3D in vitro model. Healthy cornea stromal cells (HCFs) and KC cornea stromal cells (HKCs), both male and female, were stimulated with various concentrations of E1 and E3.
Significant changes were observed between cell types, as well as between males and females in the sex hormone receptors tested; androgen receptor (AR), progesterone receptor (PR), estrogen receptor alpha (ERα), and estrogen receptor beta (ERβ) using Western blot analysis. E1 and E3 stimulations in HCF females showed AR, PR, and ERβ were significantly upregulated compared to HCF males. In contrast, ERα and ERβ had significantly higher expression in HKC’s females than HKC’s males. Our data suggest that the human cornea is a sex-dependent, hormone-responsive tissue that is significantly influenced by E1 and E3. Therefore, it is plausible that E1, E3, and sex hormone receptors are involved in the KC pathobiology, warranting further investigation.

Algorithms with Area under the Curve for Daily Urinary Estrone-3-Glucuronide and Pregnanediol-3-Glucuronide to Signal the Transition to the Luteal Phase

Background and Objectives: Home fertility assessment methods (FAMs) for natural family planning (NFP) have technically evolved with the objective metrics of urinary luteinizing hormone (LH), estrone-3-glucuronide (E3G) and pregnanediol-3-glucuronide (PDG). Practical and reliable algorithms for timing the phase of cycle based upon E3G and PDG levels are mostly unpublished and still lacking. Materials and Methods: A novel formulation to signal the transition to the luteal phase was discovered, tested, and developed with a data set of daily E3G and PDG levels from 25 women, 78 cycles, indexed to putative ovulation (day after the urinary LH surge), Day 0. The algorithm is based upon a daily relative progressive change in the ratio, E3G-AUC/PDG-AUC, where E3G-AUC and PDG-AUC are the area under the curve for E3G and PDG, respectively.
To improve accuracy the algorithm incorporated a three-fold cycle-specific increase of PDG. Results: An extended negative change in E3G-AUC/PDG-AUC of at least nine consecutive days provided a strong signal for timing the luteal phase. The algorithm correctly identified the luteal transition interval in 78/78 cycles and predicted the start day of the safe period as: Day + 2 in 10/78 cycles, Day + 3 in 21/78 cycles, Day + 4 in 28/78 cycles, Day + 5 in 15/78 cycles, and Day + 6 in 4/78 cycles. The mean number of safe luteal days with this algorithm was 10.3 ± 1.3 (SD). Conclusions: An algorithm based upon the ratio of the area under the curve for daily E3G and PDG levels along with a relative PDG increase offers another approach to time the phase of cycle. This may have applications for NFP/FAMs and clinical evaluation of ovarian function.

The 3-oxoacyl-(acyl-carrier-protein) reductase HSD-X1 of Pseudomonas citronellolis SJTE-3 catalyzes the conversion of 17β-estradiol to estrone

Background: Pseudomonas citronellolis SJTE-3 can efficiently degrade 17β-estradiol (E2) and other estrogenic chemicals. However, the enzyme responsible for E2 metabolism within strain SJTE-3 has remained unidentified.
Objective: Here, a novel 3-oxoacyl-(acyl-carrier protein) (ACP) reductase, HSD-X1 (WP_009617962.1), was identified in SJTE-3 and its enzymatic characteristics for the transformation of E2 were investigated.
Methods: Multiple sequence alignment and homology modelling were used to predict the protein structure of HSD-X1. The concentrations of different steroids in the culture of recombinant strains expressing HSD-X1 were determined by high performance liquid chromatography. Additionally, the transcription of hsd-x1 gene was investigated using reverse transcription and quantitative PCR analysis. Heterologous expression and affinity purification were used to obtain recombinant HSD-X1.
Results: The transcription of hsd-x1 gene in P. citronellolis SJTE-3 was induced by E2. Multiple sequence alignment (MSA) indicated that HSD-X1 contained the two consensus regions and conserved residues of short-chain dehydrogenase/reductases (SDRs) and 17β-hydroxysteroid dehydrogenases (17β-HSDs). Over-expression of hsd-x1 gene allowed the recombinant strain to degrade E2. Recombinant HSD-X1 was purified with a yield of 22.15 mg/L and used NAD+ as its cofactor to catalyze the oxidization of E2 into estrone (E1), while exhibiting a Km value of 0.025 ± 0.044 mM and a Vmax value of 4.92 ± 0.31 mM/min/mg. HSD-X1 could tolerate a wide range of temperature and pH, while the presence of divalent ions exerted little influence on its activity. Further, the transformation efficiency of E2 into E1 was over 98.03% across 15 min.
Conclusion: Protein HSD-X1 efficiently catalyzed the oxidization of E2 and participated in estrogen degradation by P. citronellolis SJTE-3.

Estrone(Estrone) ELISA Kit

EKF57997-48T Biomatik Corporation 48T 396.9 EUR

Estrone(Estrone) ELISA Kit

EKF57997-5x96T Biomatik Corporation 5x96T 2693.25 EUR

Estrone(Estrone) ELISA Kit

EKF57997-96T Biomatik Corporation 96T 567 EUR

Estrone(Estrone) ELISA Kit

EU3107 FN Test 96T 628.92 EUR

Rat Estrone(Estrone) ELISA Kit

ER1506 FN Test 96T 628.92 EUR

Estrone

A8426-10000 ApexBio 10g 85 EUR

Estrone

A8426-5.1 ApexBio 10 mM (in 1mL DMSO) 44 EUR

Estrone

A8426-50 ApexBio 50 mg 157.2 EUR

Estrone

A8426-5000 ApexBio 5g 62 EUR

Estrone

300149 MedKoo Biosciences 500.0mg 90 EUR

Estrone

E02400 Pfaltz & Bauer 400MG 398 EUR

Estrone

E889050 Toronto Research Chemicals 10g 91 EUR

Estrone

DE4174 Demeditec Diagnostics 96 146 EUR

Estrone

AT185 Unibiotest 1mg 1336.8 EUR

Estrone

AG185 Unibiotest 1 mg 627.6 EUR

Estrone

HY-B0234 MedChemExpress 10mM/1mL 135.6 EUR

Estrone

RM1609-1G EWC Diagnostics 1 unit 34.26 EUR

Estrone

T1009-10mg TargetMol Chemicals 10mg Ask for price

Estrone

T1009-1g TargetMol Chemicals 1g Ask for price

Estrone

T1009-1mg TargetMol Chemicals 1mg Ask for price

Estrone

T1009-50mg TargetMol Chemicals 50mg Ask for price

Estrone

T1009-5mg TargetMol Chemicals 5mg Ask for price

Estrone

TBZ2970 ChemNorm 25mg Ask for price

Estrone ELISA Kit| Rat Estrone ELISA Kit

EF018156 Lifescience Market 96 Tests 826.8 EUR

Estrone ELISA Kit| General Estrone ELISA Kit

EF019603 Lifescience Market 96 Tests 826.8 EUR

Estrone-(13,14,15,16,17,18-13C6)

E889049 Toronto Research Chemicals 25mg 21000 EUR

Temporal compositional shifts in an activated sludge microbiome during estrone biodegradation

  • Microbial biodegradation is a key process for the removal of estrogens during wastewater treatment. At least four degradation pathways for natural estrogens have been proposed. However, major estrogen degraders and the occurrence of different estrogen biodegradation pathways in wastewater treatment plants have been rarely investigated. This study was conducted to elucidate estrone biodegradation pathway and to identify key estrone-degrading bacteria in activated sludge from a major wastewater treatment plant in Bahrain. The biodegradation experiments were performed in activated sludge microcosms supplemented with estrone. Sludge samples were retrieved at time intervals to analyze the biodegradation metabolites and the temporal shifts in the bacterial community composition.
  • Chemical analysis revealed the biodegradation of more than 90% of the added estrone within 6 days, and the compounds 4-hydroxyestrone and pyridinestrone acid, which are typical markers of the 4,5-seco pathway of aerobic estrone biodegradation, were detected. Temporal shifts in the relative abundance of bacteria were most prominent among members of Proteobacteria and Bacteroidetes. While the alphaproteobacterial genera Novosphingobium and Sphingoaurantiacus were significantly enriched (from ≤ 6% to an average of 31%) in the estrone-amended activated sludge after 2 days of incubation, the bacteroidete Pedobacter was uniquely detected in these microcosms at day 10. The relative abundance of Polyangia (Nannocyctis) increased to an average of 10 ± 0.4% in the estrone-amended activated sludge after 4 days of incubation.
  • Enrichment cultivation of bacteria from the activated sludge on estrone resulted in a mixed culture that was capable of degrading estrone. An estrone-degrading strain was isolated from this mixed culture and was affiliated with the known estrogen-degrading Alphaproteobacteria Sphingobium estrogenivorans. We conclude that estrone degradation in the activated sludge from the studied wastewater treatment plant proceeds via the 4,5-seco pathway and is most likely mediated by alphaproteobacterial taxa.

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

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 42 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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  • 1 g
  • 25 g
  • 5 g

Estradiol

DE2693 Demeditec Diagnostics 96 84 EUR

Estradiol

E1KS1709 EnoGene 100 mg 151.2 EUR

Estradiol

AT070 Unibiotest 1mg 1641.6 EUR

Estradiol

AG070 Unibiotest 1 mg 627.6 EUR

Estradiol

GWB-2E7E30 GenWay Biotech 1 ml Ask for price

Estradiol

HY-B0141 MedChemExpress 500mg 129.6 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

Estradiol

GP9683-5G Glentham Life Sciences 5 g 156 EUR

Estradiol

T1048-10mg TargetMol Chemicals 10mg Ask for price

Estradiol

T1048-1g TargetMol Chemicals 1g Ask for price

Estradiol

T1048-1mg TargetMol Chemicals 1mg Ask for price

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.

Involvement of c-Jun N-terminal kinase 2 (JNK2) in Endothelin1 (ET1) Mediated Neurodegeneration of Retinal Ganglion Cells

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

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 (DHEAS) 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 DHEAS 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 DHEAS 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 ELISA

E22-HC9024 EnoGene 96T 495 EUR

DHEA-S ELISA

GWB-BQK1B7 GenWay Biotech 96 Well Plate Ask for price

DHEA-S Antibody

GWB-F09C57 GenWay Biotech 0.1 ml Ask for price

DHEA-S Standard, 350UL

C201-350UL Arbor Assays 350UL 207 EUR

DHEA-S Standard, 70UL

C201-70UL Arbor Assays 70UL 85 EUR

DHEA-S ELISA KIT|Human

EF006758 Lifescience Market 96 Tests 826.8 EUR

DHEA-S (human) ELISA Kit

K7428-100 Biovision each 940.8 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

DetectX® DHEA-S Antibody, 3ML

C199-3ML Arbor Assays 3ML 218 EUR

DetectX® DHEA-S Antibody, 13ML

C199-13ML Arbor Assays 13ML 817 EUR

DetectX® DHEA-S Conjugate, 3ML

C200-3ML Arbor Assays 3ML 218 EUR

Dehydroepiandrosterone Sulfate (DHEA-S) Antibody

abx342013-100g Abbexa 100 µg Ask for price

Dehydroepiandrosterone Sulfate (DHEA-S) Antibody

abx342013-20g Abbexa 20 µg 1162.5 EUR

Dehydroepiandrosterone Sulfate (DHEA-S) Antibody

abx342013-50g Abbexa 50 µg 4337.5 EUR

Dehydroepiandrosterone Sulphate (DHEA-S) Antibody

abx342172-100g Abbexa 100 µg Ask for price

Dehydroepiandrosterone Sulphate (DHEA-S) Antibody

abx342172-20g Abbexa 20 µg 700 EUR

Dehydroepiandrosterone Sulphate (DHEA-S) Antibody

abx342172-50g Abbexa 50 µg 2137.5 EUR

DetectX® DHEA-S Conjugate, 13ML

C200-13ML Arbor Assays 13ML 817 EUR

OKAU00048-1PLATE - DHEA-S ELISA Kit

OKAU00048-1PLATE Aviva Systems Biology 1plate 400 EUR

OKAU00048-5PLATE - DHEA-S ELISA Kit

OKAU00048-5PLATE Aviva Systems Biology 5plate 1529 EUR

DHEA

2172-100 Biovision each 130.8 EUR

DHEA

2172-500 Biovision each 301.2 EUR

DHEA

DEH3344 Demeditec Diagnostics 96 107 EUR

DHEA

HY-14650 MedChemExpress 500mg 325.2 EUR

Dehydroepiandrosterone Sulfate (DHEA-S) CLIA Kit

abx196899-100l Abbexa 100 µl Ask for price

Comparative profiling of prenatal cortisol and DHEAS 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

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.

Cyclic GMP Antibody

abx022640-02ml Abbexa 0.2 ml 1128 EUR

Cyclic GMP Antibody

GWB-BA5CC2 GenWay Biotech 0.2 ml Ask for price

Cyclic GMP Antibody

GWB-DFA25F GenWay Biotech 2 ml Ask for price

Cyclic GMP EIA Kit

SKT-211-96 Stressmarq 1 plate of 96 wells 568.8 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

ELISA kit for cGMP (Cyclic GMP)

E-EL-0083 Elabscience Biotech 1 plate of 96 wells 452.4 EUR

DetectX® Cyclic GMP Antibody, 3ML

C078-3ML Arbor Assays 3ML 218 EUR

DetectX® Cyclic GMP Antibody, 3ML

C237-3ML Arbor Assays 3ML 254 EUR

DetectX® Cyclic GMP Antibody, 13ML

C078-13ML Arbor Assays 13ML 1021 EUR

DetectX® Cyclic GMP Conjugate, 3ML

C079-3ML Arbor Assays 3ML 218 EUR

DetectX® Cyclic GMP Antibody, 13ML

C237-13ML Arbor Assays 13ML 1021 EUR

DetectX® Cyclic GMP Conjugate, 13ML

C079-13ML Arbor Assays 13ML 1021 EUR

Cyclic di-GMP

B7839-.5 ApexBio 500 µg 105 EUR

Cyclic di-GMP

B7839-1 ApexBio 1 mg 201 EUR

Human Cyclic GMP-AMP synthase (MB21D1)

1-CSB-EP822726HU Cusabio
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  • 100ug
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  • 1MG
  • 200ug
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  • 50ug

Cyclic GMP CLIA Kit (High-Sensitivity)

SKT-210-96 Stressmarq 1 plate of 96 wells 624 EUR

Cyclic GMP-AMP synthase (CGAS) Antibody

abx128130-100l Abbexa 100 µl 287.5 EUR

Cyclic GMP-AMP synthase (CGAS) Antibody

abx128130-1ml Abbexa 1 ml 825 EUR

Cyclic GMP-AMP synthase (CGAS) Antibody

abx128130-200l Abbexa 200 µl 362.5 EUR

Cyclic-di-GMP

B2045-1000 Biovision each 666 EUR

Cyclic-di-GMP

B2045-250 Biovision each 248.4 EUR

Cyclic-di-GMP

C987353 Toronto Research Chemicals 75mg 19000 EUR

Cyclic-di-GMP

HY-107780 MedChemExpress 10mM/1mL 4454.4 EUR

cyclic-di-GMP

NU-951L Jena Bioscience GmbH 5 x 1µmol 505.6 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

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

SKT-200-192 Stressmarq 2 plates of 96 wells 226 EUR

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

NGAL (Detection Ab)

abx019242-100ug Abbexa 100 ug 777.6 EUR

SEB Detection Kit

6030 Chondrex 1 kit 377 EUR

LPS Detection Kit

6039 Chondrex 1 kit 404 EUR

TLR Detection Set

PSI-1806 ProSci 1 Set 1627.8 EUR

PD1 Detection Set

SD8600 ProSci 1 Set 537.9 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

HROS Detection Kit

FLAPF100-2 Cell Technology 150 Tests 280 EUR

IRAK Detection Set

PSI-1802 ProSci 1 Set 884.4 EUR

ORAI Detection Set

PSI-1819 ProSci 1 Set 1627.8 EUR

Grik Detection Set

PSI-1824 ProSci 1 Set 1627.8 EUR

PDL1 Detection Set

SD8500 ProSci 1 Set 355.2 EUR

LAG3 Detection Set

SD8700 ProSci 1 Set 569.4 EUR

Cell Detection Dye

CDD200 ProFoldin 0.1 ml 153.27 EUR

Human Urinary oxalate(urinary oxalate) ELISA Kit

QY-E05414 Qayee Biotechnology 96T 433.2 EUR

4x Detection Buffer

79389 BPS Bioscience 10 ml 195 EUR

OVA Detection ELISA

6032 Chondrex 1 kit 377 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

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 Stressmarq 2 plates of 96 wells 248 EUR

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

B1717-1000 ApexBio 1g 34 EUR

Creatinine

B1717-50 ApexBio 50 mg 153.6 EUR

Creatinine

B1717-5000 ApexBio 5g 48 EUR

Creatinine

C271-100MG TOKU-E 100 mg 223.26 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.

Extension of Genetic Marker List Using Unnatural Amino Acid System: An Efficient Genomic Modification Strategy in Escherichia coli.

Extension of Genetic Marker List Using Unnatural Amino Acid System: An Efficient Genomic Modification Strategy in Escherichia coli.
Genetic manipulations together with chromosome engineering are important strategies used to restructure cell metabolism. Lambda/Red (λ/Red)-mediated recombination is probably the most generally utilized strategy for chromosomal modulation in Escherichia coli. However, the effectivity of this technique is considerably hampered by the laborious elimination of the selectable markers.
To overcome the issue, the combination helper plasmid was constructed, pSBC1a-CtR, which comprises Red recombinase, Cre recombinase, and exogenous orthogonal aminoacyl-transfer RNA (tRNA) synthetase/tRNA pairs, permits an unnatural amino acid (UAA) to be genetically encoded on the outlined website of the antibiotic resistance gene-encoded protein. When UAAs are usually not in the tradition medium, there was no expression in the antibiotic resistance gene-encoded protein. Accordingly, the following process of antibiotic gene excising is just not wanted.
To confirm this technique, poxB gene was knocked out efficiently. Furthermore, sequential deletion of three goal genes (galR, ptsG, and pgi) was in a position to generate neurosporene-producing pressure marked by excessive development fee. Thus, the site-specific incorporation UAA mutagenesis system have been used to regulate and broaden the use of conditional selectable marker, and the approach is used to facilitate a speedy steady genome modifying in Escherichia coli.