Cell-type specific interaction of endothelin and the nitric oxide system: pattern of prepro-ET–1 expression in kidneys of L-NAME treated prepro-ET–1 promoter-lacZ-transgenic mice.

by
  • Nitric oxide (NO) and endothelin-1 (ET-1) are known to play a major role in renal and vascular pathophysiology and exhibit a close interaction with ET-1, stimulating NO production; NO in turn inhibits ET-1 expression. Our objectives were (1) to establish a novel transgenic mouse model facilitating ET-1 expression assessment in vivo, (2) to validate this model by assessing prepro-ET-1 promoter activity in mice embryos by means of our novel model and comparing expression sites to well-established data on ET-1 in fetal development and (3) to investigate renal ET-NO interaction by assessing prepro-ET-1 promoter activity in different structures of the renal cortex in the setting of blocked NO synthases via L-NAME administration. We established transgenic mice carrying a lacZ reporter gene under control of the human prepro-ET-1 gene promoter sequence (8 kb of 5′ sequences).
  • Bluo-Gal staining of tissue sections revealed intracellular blue particles as indicators of prepro-ET-1 promoter activity. In mouse embryos, we detected high prepro-ET-1 promoter activity in the craniofacial region, as well as in bone and cartilage consistent with the literature. In order to investigate the interaction of ET-1 and NO in the kidney in vivo, transgenic mice at the age of 3-4 months were treated with a single dose of the NO synthase inhibitor L-NAME (25 mg (kg bw)(-1) i.p.) 12 h before kidney removal. Bluo-Gal staining of kidney sections revealed intracellular blue particles as indicators of prepro-ET-1 promoter activity in tubular and vascular endothelium and glomerular cells.
  • Particle count was closely correlated to kidney tissue ET-1 content (R=0.918, P<0.001). Comparison of counts revealed an increase by 135+/-53% in L-NAME treated (n=12) compared to non-treated mice (n=10, P=0.001). Cell-type specific evaluation revealed an increase of 136+/-51% in tubular (P=0.001) and 105+/-41% in glomerular cells (P=0.046), but no significant increase in vascular endothelium. In conclusion, our study revealed a close interaction of renal endothelin and the NO system in a cell-type specific manner. Our new transgenic model provides a unique opportunity to analyse regulation of the ET system on a cellular level in vivo.

<em>Endothelin</em>-<em>1</em> promotes cell survival in renal cell carcinoma through the <em>ET</em>(A) receptor.

Endothelin-1 (ET-1) is a potent vasoconstrictor that has been shown to significantly impact many benign and malignant tissues by signaling through its two cognate receptors: ET(A) and ET(B). As ET-1 has a role in both normal and diseased kidney, we initiated studies to investigate endothelin axis expression and function in renal cell carcinoma (RCC). In this study, relatively high levels of ET-1 were detected in all six human RCC cell lines investigated. RT-PCR and Southern analyses revealed that all six RCC cell lines expressed ET(A) receptor mRNA, while 3/6 cell lines also expressed ET(B) mRNA.
High affinity ET-1 binding occurred in all but one RCC cell line and quantitative RT-PCR demonstrated ET(A) mRNA expression in all six cell lines. Methylation of the ET(B) promoter (EDNRB) in 4/6 RCC cell lines was observed, suggesting a mechanism for repressed ET(B) expression. Moreover, methylation occurred in 32/48 of renal tumors and in 27/55 of histologically normal adjacent tissue samples studied, while no methylation was evident in any normal tissue isolated from nephrectomy or at autopsy. Functionally, ET-1 significantly inhibited paclitaxel-induced apoptosis in RCC cells through binding ET(A) with the ET-1 signaling mediated via the PI3-kinase/Akt pathway. Collectively, these data support the therapeutic targeting of the ET(A) receptor as a novel treatment strategy for RCC.

<em>Endothelin</em>-<em>1</em> and its receptors <em>ET</em>(A) and <em>ET</em>(B) in drug-induced gingival overgrowth.

BACKGROUND
The purpose of this study was to study the expression of endothelin-1 (ET-1) and its receptors ETA and ETB in normal human gingiva and cyclosporin-induced gingival fibroblasts.
METHODS
Gingival samples were collected from eight normal healthy individuals, eight patients with periodontitis, and eight patients with cyclosporin A (CsA)-induced gingival overgrowth. Total RNA was extracted from tissue samples, and reverse transcriptase-polymerase chain reaction was performed for ET-1, ETA, and ETB. ET-1 protein was estimated from the tissues by enzyme-linked immunosorbent assay. The expression of ET-1 and its receptors was also examined in gingival fibroblast cells treated with CsA.
RESULTS
ET-1 mRNA expression was significantly higher in patients with CsA-induced gingival overgrowth (P <0.001) than in patients with periodontitis and the controls. ETA mRNA was expressed more than the ETB in all examined samples. In human gingival fibroblasts, ET-1 expression was increased with CsA incorporation compared to controls (P <0.001).
CONCLUSIONS
These results suggest that CsA can modulate the expression of ET-1 in gingival fibroblasts and CsA-induced gingival overgrowth.

Regulation and expression of <em>endothelin</em>-<em>1</em> (<em>ET</em>-<em>1</em>) and <em>ET</em>-receptors in rat epithelial cells of renal and intestinal origin.

  • The hormone endothelin-1 (ET-1) is involved in many functions of the kidney and intestine. In addition to its vasoactive and proliferative effects, ET-1 is involved in the maintenance of water and salt balance, and in drug excretion by influencing the activity of different transporters in the epithelial cells of these two organs. To study ET-1 function and its role in pathophysiological processes in epithelial cells in vitro, we investigated ET-1 and ET-receptor expression and inducibility of ET-1 excretion by cytokines in three rat cell lines of intestinal (IEC-6) and renal (NRK-52E and GERP) origin.
  • Immunocytochemistry showed that all three cell lines express ET-1 and the ET-A and ET-B receptor. ET-1 was expressed intracellularly, and also the ET-A receptor showed a punctate intracellular staining pattern. The ET-B receptor was localized in the membrane, which was confirmed by Western blot analysis. Real-time RT-PCR and ELISA showed that exposure of IEC-6 cells to the cytokines, interleukin-1beta (IL-1beta) and tumor necrosis factor-alpha (TNFalpha), induced ET-1 mRNA expression and excretion, while IL-2 was ineffective.
  • In NRK-52E cells, IL-1beta and TNFalpha induced ET-1 excretion as well. In GERP cells, adequate measurement of cytokine effects on ET-1 excretion was not possible, since ET-1 excretion under non-stimulated conditions was around the lowest level of detection. In conclusion, we showed ET-1 and ET-receptor expression, and inducibility of ET-1 by cytokines in IEC-6, NRK-52E, and GERP cells.
  • These rat intestinal and renal cell lines appear to be suitable for further characterisation of ET-1 function and its role in pathophysiological processes in epithelial cells.

Endothelin 1 (ET-1) Antibody
abx024086-100ug Abbexa 100 ug 1128 EUR

Endothelin-1 (ET-1), big (Rat)
023-32 PHOENIX PEPTIDE 100 μg 206.28 EUR

Endothelin-1 (ET-1), big (Human)
023-10 PHOENIX PEPTIDE 100 μg 154.44 EUR

ET-1(Endothelin-1) ELISA Kit
EKF60092-48T Biomatik Corporation 48T 396.9 EUR

ET-1(Endothelin-1) ELISA Kit
EKF60092-5x96T Biomatik Corporation 5x96T 2693.25 EUR

ET-1(Endothelin-1) ELISA Kit
EKF60092-96T Biomatik Corporation 96T 567 EUR

ET-1(Endothelin-1) ELISA Kit
EU0205 FN Test 96T 628.92 EUR

ET-1/Endothelin 1 Rabbit pAb
E2252197 EnoGene 100ul 225 EUR

Endothelin, pan (Endothelin 1, Endothelin-1, EDN1, ET1, ET-1, Endothelin 2, Endothelin-2, EDN2, ET2, ET-2, Endothelin 3, Endothelin-3, EDN3, ET3, ET-3, HDLCQ7, MGC15067, MGC61498, Preproendothelin-1, PPET1, Preproendothelin-2, PPET2, Preproendothelin-3, P
MBS6250834-01mL MyBiosource 0.1(mL 865 EUR

Endothelin, pan (Endothelin 1, Endothelin-1, EDN1, ET1, ET-1, Endothelin 2, Endothelin-2, EDN2, ET2, ET-2, Endothelin 3, Endothelin-3, EDN3, ET3, ET-3, HDLCQ7, MGC15067, MGC61498, Preproendothelin-1, PPET1, Preproendothelin-2, PPET2, Preproendothelin-3, P
MBS6250834-5x01mL MyBiosource 5x0.1mL 3750 EUR

Endothelin, pan (Endothelin 1, Endothelin-1, EDN1, ET1, ET-1, Endothelin 2, Endothelin-2, EDN2, ET2, ET-2, Endothelin 3, Endothelin-3, EDN3, ET3, ET-3, HDLCQ7, MGC15067, MGC61498, Preproendothelin-1, PPET1, Preproendothelin-2, PPET2, Preproendothelin-3, P
MBS6247892-01mL MyBiosource 0.1(mL 865 EUR

Endothelin, pan (Endothelin 1, Endothelin-1, EDN1, ET1, ET-1, Endothelin 2, Endothelin-2, EDN2, ET2, ET-2, Endothelin 3, Endothelin-3, EDN3, ET3, ET-3, HDLCQ7, MGC15067, MGC61498, Preproendothelin-1, PPET1, Preproendothelin-2, PPET2, Preproendothelin-3, P
MBS6247892-5x01mL MyBiosource 5x0.1mL 3750 EUR

Endothelin, pan (Endothelin 1, Endothelin-1, EDN1, ET1, ET-1, Endothelin 2, Endothelin-2, EDN2, ET2, ET-2, Endothelin 3, Endothelin-3, EDN3, ET3, ET-3, HDLCQ7, MGC15067, MGC61498, Preproendothelin-1, PPET1, Preproendothelin-2, PPET2, Preproendothelin-3, P
MBS6248913-01mL MyBiosource 0.1(mL 865 EUR

Endothelin, pan (Endothelin 1, Endothelin-1, EDN1, ET1, ET-1, Endothelin 2, Endothelin-2, EDN2, ET2, ET-2, Endothelin 3, Endothelin-3, EDN3, ET3, ET-3, HDLCQ7, MGC15067, MGC61498, Preproendothelin-1, PPET1, Preproendothelin-2, PPET2, Preproendothelin-3, P
MBS6248913-5x01mL MyBiosource 5x0.1mL 3750 EUR

Endothelin, pan (Endothelin 1, Endothelin-1, EDN1, ET1, ET-1, Endothelin 2, Endothelin-2, EDN2, ET2, ET-2, Endothelin 3, Endothelin-3, EDN3, ET3, ET-3, HDLCQ7, MGC15067, MGC61498, Preproendothelin-1, PPET1, Preproendothelin-2, PPET2, Preproendothelin-3, P
MBS6249840-01mL MyBiosource 0.1(mL 865 EUR

Endothelin, pan (Endothelin 1, Endothelin-1, EDN1, ET1, ET-1, Endothelin 2, Endothelin-2, EDN2, ET2, ET-2, Endothelin 3, Endothelin-3, EDN3, ET3, ET-3, HDLCQ7, MGC15067, MGC61498, Preproendothelin-1, PPET1, Preproendothelin-2, PPET2, Preproendothelin-3, P
MBS6249840-5x01mL MyBiosource 5x0.1mL 3750 EUR

Endothelin, pan (Endothelin 1, Endothelin-1, EDN1, ET1, ET-1, Endothelin 2, Endothelin-2, EDN2, ET2, ET-2, Endothelin 3, Endothelin-3, EDN3, ET3, ET-3, HDLCQ7, MGC15067, MGC61498, Preproendothelin-1, PPET1, Preproendothelin-2, PPET2, Preproendothelin-3, P
MBS6251787-01mL MyBiosource 0.1(mL 865 EUR

Endothelin, pan (Endothelin 1, Endothelin-1, EDN1, ET1, ET-1, Endothelin 2, Endothelin-2, EDN2, ET2, ET-2, Endothelin 3, Endothelin-3, EDN3, ET3, ET-3, HDLCQ7, MGC15067, MGC61498, Preproendothelin-1, PPET1, Preproendothelin-2, PPET2, Preproendothelin-3, P
MBS6251787-5x01mL MyBiosource 5x0.1mL 3750 EUR

Endothelin, pan (Endothelin 1, Endothelin-1, EDN1, ET1, ET-1, Endothelin 2, Endothelin-2, EDN2, ET2, ET-2, Endothelin 3, Endothelin-3, EDN3, ET3, ET-3, HDLCQ7, MGC15067, MGC61498, Preproendothelin-1, PPET1, Preproendothelin-2, PPET2, Preproendothelin-3, P
MBS6254880-01mL MyBiosource 0.1(mL 865 EUR

Endothelin, pan (Endothelin 1, Endothelin-1, EDN1, ET1, ET-1, Endothelin 2, Endothelin-2, EDN2, ET2, ET-2, Endothelin 3, Endothelin-3, EDN3, ET3, ET-3, HDLCQ7, MGC15067, MGC61498, Preproendothelin-1, PPET1, Preproendothelin-2, PPET2, Preproendothelin-3, P
MBS6254880-5x01mL MyBiosource 5x0.1mL 3750 EUR

Endothelin, pan (Endothelin 1, Endothelin-1, EDN1, ET1, ET-1, Endothelin 2, Endothelin-2, EDN2, ET2, ET-2, Endothelin 3, Endothelin-3, EDN3, ET3, ET-3, HDLCQ7, MGC15067, MGC61498, Preproendothelin-1, PPET1, Preproendothelin-2, PPET2, Preproendothelin-3, P
MBS6253840-01mL MyBiosource 0.1(mL 865 EUR

Endothelin, pan (Endothelin 1, Endothelin-1, EDN1, ET1, ET-1, Endothelin 2, Endothelin-2, EDN2, ET2, ET-2, Endothelin 3, Endothelin-3, EDN3, ET3, ET-3, HDLCQ7, MGC15067, MGC61498, Preproendothelin-1, PPET1, Preproendothelin-2, PPET2, Preproendothelin-3, P
MBS6253840-5x01mL MyBiosource 5x0.1mL 3750 EUR

Endothelin, pan (Endothelin 1, Endothelin-1, EDN1, ET1, ET-1, Endothelin 2, Endothelin-2, EDN2, ET2, ET-2, Endothelin 3, Endothelin-3, EDN3, ET3, ET-3, HDLCQ7, MGC15067, MGC61498, Preproendothelin-1, PPET1, Preproendothelin-2, PPET2, Preproendothelin-3, P
MBS6252800-01mL MyBiosource 0.1(mL 865 EUR

Endothelin, pan (Endothelin 1, Endothelin-1, EDN1, ET1, ET-1, Endothelin 2, Endothelin-2, EDN2, ET2, ET-2, Endothelin 3, Endothelin-3, EDN3, ET3, ET-3, HDLCQ7, MGC15067, MGC61498, Preproendothelin-1, PPET1, Preproendothelin-2, PPET2, Preproendothelin-3, P
MBS6252800-5x01mL MyBiosource 5x0.1mL 3750 EUR

Endothelin, pan (Endothelin 1, Endothelin-1, EDN1, ET1, ET-1, Endothelin 2, Endothelin-2, EDN2, ET2, ET-2, Endothelin 3, Endothelin-3, EDN3, ET3, ET-3, HDLCQ7, MGC15067, MGC61498, Preproendothelin-1, PPET1, Preproendothelin-2, PPET2, Preproendothelin-3, P
MBS6256959-01mL MyBiosource 0.1mL 865 EUR

Endothelin, pan (Endothelin 1, Endothelin-1, EDN1, ET1, ET-1, Endothelin 2, Endothelin-2, EDN2, ET2, ET-2, Endothelin 3, Endothelin-3, EDN3, ET3, ET-3, HDLCQ7, MGC15067, MGC61498, Preproendothelin-1, PPET1, Preproendothelin-2, PPET2, Preproendothelin-3, P
MBS6256959-5x01mL MyBiosource 5x0.1mL 3750 EUR

[Relationship of <em>endothelin</em>-<em>1</em> (<em>ET</em>-<em>1</em>) TaqI and tumor necrosis factor (TNF) a gene polymorphism with portal hypertension in liver cirrhosis].

OBJECTIVE
To study whether liver cirrhosis and portal hypertension are associated with ET-1 TaqI polymorphism and TNFa promoter-308G to A polymorphism.
METHODS
A case control study of 106 patients with liver cirrhosis following HBV C infection was performed in comparison with 108 controls by PCR-RFLP.
RESULTS
The frequency of C allele and CC+TC genotype in TaqI polymorphism of ET-1 gene in the portal hypertension group (LC+) was significantly higher than that in the healthy controls, and the frequency of TNF2/1 genotype in TNFa promoter -308 G to A polymorphism in LC+ group was significantly higher than that in the control group. The results by stratification analysis showed that TCF2 genotype frequency was higher in the LC+ group than in the control group. ET-1 TaqI polymorphism and TNFa polymorphism were risk factors for the occurrence of portal hypertension by Logistic regression analysis.
CONCLUSIONS
ET-1 TaqI polymorphism and TNFa polymorphism are associated with portal hypertension, and are new risk factors for the occurrence of portal hypertension. TCF2 genotype may be a susceptible gene of portal hypertension.

Comparative Analysis between Urinary Calprotectin and Serum Creatinine for Early Detection of Intrinsic Acute Kidney Injury

by
Background: Acute kidney injury (AKI) is a common and important clinical condition that may lead to chronic kidney disease if it is not diagnosed and treated in its early stages. Urinary calprotectin is a valuable recognized biomarker that can be used to differentiate prerenal and intrinsic AKI. However, till date only a few reports on urine calprotectin measurement in early diagnosis of intrinsic AKI are available. In this study, we compared the sensitivity and specificity of urinary calprotectin with those of serum creatinine in detecting early intrinsic AKI.
Methods: Over 6 months period (April to October 2018), 81 of 408 patients admitted to the pediatric intensive care unit met the criteria of this cross-sectional study. Their serum creatinine and urinary calprotectin were measured on the first and third day of admission using Jaffe and Elisa radioimmunoassay methods, respectively. The AKI was defined according to the pRIFLE criteria.
Results: Of the total 81 patients, 67 had the criteria of intrinsic AKI. Of these 62% were female and 38% were male. The mean age of the patients was 22 months. According to data analysis, the area under the curve of ROC of urinary calprotectin on day-1 to detect renal failure is 0.93 with the best cutoff point obtained at 530 ng/mL. The sensitivity, specificity, positive, and negative predictive values of urinary calprotectin levels in diagnosing AKI at this cutoff point are 92.5%, 92.8%, 98.4, and 72.2%, respectively. Besides, urinary calprotectin changes occur much earlier than the rising of serum creatinine.
Conclusion: Urinary level of calprotectin is a very sensitive biomarker for early diagnosis of intrinsic AKI in children and it can be used in intensive care units or anywhere critically ill children admitted to detect intrinsic AKI. Besides, this study shows that urine calprotectin may be a more sensitive and specific biomarker than serum creatinine in the early phases of intrinsic AKI.

High-performance surface-enhanced Raman spectroscopy chip integrated with a micro-optical system for the rapid detection of creatinine in serum

To improve the sensitivity of disease biomarker detection, we proposed a high-performance surface-enhanced Raman spectroscopy (SERS) chip integrated with a micro-optical system (MOS). The MOS, which is based on the micro-reflecting cavity and the micro-lens, optimizes the optical matching characteristics of the SERS substrate and the Raman detection system, and greatly improves the SERS detection sensitivity by improving the collection efficiency of the Raman scattering signal. A uniform single layer of silver nanoparticles on a gold film was prepared as the SERS substrate using a liquid-liquid interface self-assembly method. The micro-reflecting cavity and micro-lens were prepared using micro-processing technology. The SERS chip was constructed based on the MOS and the Au film-based SERS substrate, and experimental results showed an EF of 1.46×108, which is about 22.4 times higher than that of the Si-based SERS substrate.
The chip was used for the detection of creatinine and the detection limit of creatinine in aqueous solution was 1 µM while the detection limit in serum was 5 µM. In addition, SERS testing was conducted on serum samples from normal people and patients with chronic renal impairment. Principal component analysis and linear discriminant analysis were used for modeling and identification, and the results showed a 90% accuracy of blind sample detection. These results demonstrate the value of this SERS chip for both research and practical applications in the fields of disease diagnosis and screening.

Drain fluid creatinine-to-serum creatinine ratio as an initial test to detect urine leakage following cystectomy: A retrospective study

Introduction: Urine leak following radical cystectomy is a known complication. Among the various methods to diagnose this, assessment of drain fluid creatinine is a relatively easy procedure. We aimed to ascertain the validity of the drain fluid creatinine-to-serum creatinine ratio (DCSCR) as an initial indicator of urinary leak in patients undergoing radical cystectomy.
Methods: We retrospectively identified consecutive patients with documentation of drain fluid creatinine in the postoperative period following cystectomy and urinary diversion at our institution between January 2009 and December 2018. All continent diversions and any patient with a DCSCR >1.5:1 underwent contrast study postoperatively. A diagnosis of urine leak was made following confirmatory imaging. Receiver operative characteristic curves were created, and Youden’s index was used to determine the strength and clinical utility of DCSCR as a diagnostic test.
Results: Two hundred forty-four of the 340 patients included in the study underwent cystectomy with conduit and 81 underwent neobladder creation. Sixteen out of 340 (4.7%) patients had radiologically confirmed urinary leak. DCSCR was elevated in all ureteric anastomotic leaks and in 1 out of the 7 neobladder-urethral anastomotic (NUA) leaks. The sensitivity and specificity of DCSCR to predict all urinary leaks were 68.8% and 80.9% at 1.12 (area under the curve [AUC] = 0.838), whereas at a value of 1.18 (AUC = 0.876) and with the exclusion of NUA leaks, the sensitivity was 77.8% and specificity was 87.6%.
Conclusions: DCSCR is a good preliminary test for identifying patients who need prompt confirmatory testing for localizing urinary leaks. A drain creatinine level just 18% higher than the serum creatinine level can signify a urine leak. This is different from general assumptions of a higher DCSCR.

Creatinine Serum Detection Kit
SKT-217-192 Stressmarq 2 plates of 96 wells 186 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

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 Stressmarq 2 plates of 96 wells 169.5 EUR

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

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

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

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 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

DetectX® Creatinine Reagent, 20ML
C004-20ML Arbor Assays 20ML 254 EUR

DetectX® Creatinine Reagent, 50ML
C004-50ML Arbor Assays 50ML 360 EUR

Creatinine Serum Low Sample Volume Kit (384-well Plate)
KB02-H1D Arbor Assays 1x384 well plate 431 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
09626-34 NACALAI TESQUE 5G 11.55 EUR

Utility of measuring serum creatinine to detect renal compromise in ED patients receiving IV contrast-enhanced CT scan

Objective: The objectives of this study are to determine the efficacy of a roster of clinical factors in identifying risk for renal insufficiency in emergency department (ED) patients requiring intravenous contrast-enhanced CT scan (IVCE-CT) and to help mitigate potential for developing contrast-induced nephropathy (CIN).
Methods: A review was conducted of consecutive ED patients who received IVCE-CT during a 4-month period in our urban ED. The values of ED serum creatinine (SCr) performed were tabulated. The medical records of all patients with an elevated SCr (> 1.4 mg/dL) were reviewed to determine and correlate the presence of clinical risk factors for underlying renal insufficiency.
Results: During the 4-month study period, there were 2260 consecutive cases who received IVCE-CT; of these, 2250 (99.6%) had concomitant measurement of SCr. Elevated SCr occurred in 141 patients (6.2%); of these, 75 had a SCr > 2 mg/dL. In all, 139/141 (98.6%) with an elevated SCr had an underlying chronic or acute medical condition identified by medical record review which potentially compromised renal function, including chronic renal disease, diabetes mellitus, HIV infection, cancer, hypertension, congestive heart failure, sepsis/septic shock, chronic alcoholism, and sickle cell disease. Two patients with no identified risk factor each had (mildly) elevated SCr; both had a normal SCr measured post-CT scan. The total cost of performing serum basic metabolic panel to measure SCr in all patients during the 4-month study period was $94,500.
Conclusions: Elevated SCr is rarely present in ED patients without recognized risk factors who receive IVCE-CT scan. The vast majority with underlying renal insufficiency are readily identified by a review of the patient’s medical history and/or clinical findings. Routine SCr measurement on all ED patients regardless of risk stratification prior to IVCE imaging is neither time nor cost-effective.

Potent repression of C-reactive protein (CRP) expression by the JAK1/2 inhibitor ruxolitinib in inflammatory human hepatocytes

by
To determine whether inflammatory hepatocytes may constitute primary targets for ruxolitinib, a Janus kinase (JAK) inhibitor, its effects towards expression of hepatic acute-phase proteins, especially C-reactive protein (CRP), were assessed.Ruxolitinib effects were analysed in primary human hepatocytes and human hepatoma HepaRG cells exposed to various inflammatory stimuli.
RESULTS

Ruxolitinib was found to fully inhibit lipopolysaccharide (LPS)-induced CRP secretion and mRNA expression, at concentrations (IC50 = 12.9 nM) achievable in human blood. It similarly repressed CRP up-regulation due to several Toll-like receptor agonists or pro-inflammatory cytokines [interleukin (IL) 1β, IL6 and tumour necrosis factor α] and counteracted LPS-mediated induction of serum amyloid A, fibrinogen, haptoglobin and serpin. Ruxolitinib was additionally found to block the activation of the IL6/JAK/signal transducer and activator of transcription (STAT) pathway triggered by LPS and whose inhibition by the neutralizing anti-IL6 receptor antibody tocilizumab prevented CRP induction.Ruxolitinib can potently repress induction of CRP in inflammatory human hepatocytes, most likely through targeting the IL6/JAK/STAT signalling cascade. Hepatic production of acute-phase proteins during liver inflammation may, therefore, constitute a target for ruxolitinib.

Nanomolar aluminum induces expression of the inflammatory systemic biomarker C–reactive protein (CRP) in human brain microvessel endothelial cells (hBMECs).

C-reactive protein (CRP; also known as pentraxin 1, PTX1), a 224 amino acid soluble serum protein organized into a novel pentameric ring-shaped structure, is a highly sensitive pathogenic biomarker for systemic inflammation. High CRP levels are found in practically every known inflammatory state, and elevated CRP levels indicate an increased risk for several common age-related human degenerative disorders, including cardiovascular disease, cancer, diabetes, and Alzheimer’s disease (AD). While the majority of CRP is synthesized in the liver for secretion into the systemic circulation, it has recently been discovered that an appreciable amount of CRP is synthesized in highly specialized endothelial cells that line the vasculature of the brain and central nervous system (CNS).
These highly specialized cells, the major cell type lining the human CNS vasculature, are known as human brain microvessel endothelial cells (hBMECs). In the current pilot study we examined (i) CRP levels in human serum obtained from AD and age-matched control patients; and (ii) analyzed the effects of nanomolar aluminum sulfate on CRP expression in primary hBMECs. The three major findings in this short communication are: (i) that CRP is up-regulated in AD serum; (ii) that CRP serum levels increased in parallel with AD progression; and (iii) for the first time show that nanomolar aluminum potently up-regulates CRP expression in hBMECs to many times its ‘basal abundance’. The results suggest that aluminum-induced CRP may in part contribute to a pathophysiological state associated with a chronic systemic inflammation of the human vasculature.

High sensitivity C–reactive protein (Hs-CRP) remains highly stable in long-term archived human serum.

BACKGROUND
The stability of biomarkers in stored biomedical samples is crucial, especially when storage is for extended periods of time. High-sensitivity CRP (Hs-CRP) is a biomarker of low grade inflammation that is extensively used to identify and study cardiovascular and/or inflammatory processes in clinical care and large epidemiologic studies. Therefore, assessing Hs-CRP stability in archived samples at a given temperature is important to ensure precision of measurements over time and the validity of studies using archived samples.
METHODS
We evaluated the stability of Hs-CRP in 30 randomly selected human serum samples by measuring Hs-CRP concentrations in freshly collected sample [Hs-CRP (0)] and in the same set of samples after 7-11years of storage at -80°C [Hs-CRP (LT)].
RESULTS
Hs-CRP did not significantly change up to 11years of storage at -80°C as shown by a negligible median difference between Hs-CRP (0) and Hs-CRP (LT), delta(Hs-CRP (0)-Hs-CRP (LT))=-0.01, p=0.45. There was a good concordance and agreement between Hs-CRP (0) and Hs-CRP (LT) as measured respectively by Lin’s coefficient of correlation (ρC=0.98) and Bland-Altman analysis (mean difference=-0.02, 95% CI [-0.04-0.0045] p=0.107). In addition, the data also suggest that the time elapsed between collection and Hs-CRP measurement does not affect Hs-CRP stability over time when samples are kept under the appropriate conditions.
CONCLUSIONS
Long-term storage at -80°C for up to 11years did not significantly affect the stability of serum Hs-CRP. Given the cost and time for collecting fresh samples, this observation represents an important finding for biomedical research and clinical care.

C–reactive protein (CRP) induces chemokine secretion via CD11b/ICAM-1 interaction in human adherent monocytes.

Several studies support C-reactive protein (CRP) as a systemic cardiovascular risk factor. The recent detection of CRP in arterial intima suggests a dual activity in atherosclerosis as a circulating and tissue mediator on vascular and immune cells. In the present paper, we focused on the inflammatory effects of CRP on human monocytes, which were isolated by Ficoll-Percoll gradients and cultured in adherence to polystyrene, endothelial cell monolayer, or in suspension. Chemokine levels, adhesion molecule, and chemokine receptor expression were detected by ELISA, flow cytometry, and real-time RT-PCR. Migration assays were performed in a Boyden chamber. Stimulation with CRP induced release of CCL2, CCL3, and CCL4 in adherent monocytes through the binding to CD32a, CD32b, and CD64, whereas no effect was observed in suspension culture.
This was associated with CRP-induced up-regulation of adhesion molecules membrane-activated complex 1 (Mac-1) and ICAM-1 on adherent monocytes. Blockade of Mac-1/ICAM-1 interaction inhibited the CRP-induced chemokine secretion. In addition, CRP reduced mRNA and surface expression of corresponding chemokine receptors CCR1, CCR2, and CCR5 in adherent monocytes. This effect was a result of chemokine secretion, as coincubation with neutralizing anti-CCL2, anti-CCL3, and anti-CCL4 antibodies reversed the effect of CRP. Accordingly, a reduced migration of CRP-treated monocytes to CCL2 and CCL3 was observed. In conclusion, our data suggest an in vitro model to study CRP activities in adherent and suspension human monocytes. CRP-mediated induction of adhesion molecules and a decrease of chemokine receptors on adherent monocytes might contribute to the retention of monocytes within atherosclerotic lesions and recruitment of other circulating cells.

Human C Reactive Protein (CRP) Protein
20-abx168571 Abbexa
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Human C Reactive Protein (CRP) Protein
abx168571-1ml Abbexa 1 ml 175 EUR

Human C Reactive Protein (CRP) Protein
abx065612-100g Abbexa 100 µg 325 EUR

Human C Reactive Protein (CRP) Protein
20-abx065612 Abbexa
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Human C Reactive Protein (CRP) Protein
abx065612-10g Abbexa 10 µg 162.5 EUR

Human C Reactive Protein (CRP) Protein
abx065612-50g Abbexa 50 µg 250 EUR

C Reactive Protein (CRP)
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C Reactive Protein (CRP)
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C Reactive Protein (CRP)
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C Reactive Protein (CRP)
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C Reactive Protein (CRP)
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C Reactive Protein (CRP)
MBS590047-96Wells MyBiosource 96Wells 675 EUR

C Reactive Protein (CRP)
THP-0256 Creative BioMart 10mg 3198.4 EUR

Human C Reactive Protein (CRP) CLIA Kit
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Human C Reactive Protein (CRP) CLIA Kit
EKN50029-48T Biomatik Corporation 48T 414.89 EUR

Human C Reactive Protein (CRP) CLIA Kit
EKN50029-5x96T Biomatik Corporation 5x96T 2815.33 EUR

Human C Reactive Protein (CRP) CLIA Kit
EKN50029-96T Biomatik Corporation 96T 592.7 EUR

Human C Reactive Protein (CRP) CLIA Kit
EKU08918-48T Biomatik Corporation 48T 666.82 EUR

Human C Reactive Protein (CRP) CLIA Kit
EKU08918-5x96T Biomatik Corporation 5x96T 4524.85 EUR

Human C Reactive Protein (CRP) CLIA Kit
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Human C Reactive Protein (CRP) CLIA Kit
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Human C Reactive Protein (CRP) CLIA Kit
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Human C Reactive Protein (CRP) CLIA Kit
MBS2708864-96StripWells MyBiosource 96-Strip-Wells 480 EUR

Polymorphism in the human C–reactive protein (CRP) gene, serum concentrations of CRP, and the difference between intracranial and extracranial atherosclerosis.

BACKGROUND
C-reactive protein, a proinflammatory factor, is involved in the development of atherosclerosis. The CRP 1059G>C polymorphism appeared to be a susceptive marker for atherosclerosis. We investigated the relationship of the distribution of cerebral atherosclerosis with triggered serum CRP concentrations following acute ischemic stroke/transient ischemic attack (IS/TIA) and CRP 1059G>C polymorphism.
METHODS
We recruited 222 IS/TIA patients (122 with only intracranial atherosclerotic lesions and 100 with isolated extracranial atherosclerotic lesions) and 227 controls. Intra- and extracranial atherosclerotic lesions were determined by digital subtraction angiography. Serum CRP concentrations were measured by particle-enhanced immunonephelometry assay. CRP 1059G>C genotypes were obtained through PCR amplification and restriction enzyme digestion.
RESULTS
CRP concentrations were significantly higher in intra- and extracranial groups than in controls. No significant difference was found in CRP concentrations between intra- and extracranial groups. The CRP 1059G>C single-nucleotide polymorphism did not influence CRP serum concentrations. CRP genotype and allele frequencies did not differ significantly between patients and controls. However, the frequencies of GC genotype and C allele were significantly higher in extracranial group than that in intracranial group. The GC individuals showed a higher risk of extracranial atherosclerosis compared with GG individuals (OR 3.41; 95%CI, 1.124-10.347; P=0.030).
CONCLUSIONS
Serum CRP is associated with cerebral atherosclerotic disease. CRP 1059G>C polymorphism is one possible genetic determinant for the difference between intra- and extracranial atherosclerosis.

Telomere shortening is associated with corticosterone stress response in adult barn swallows

by
When vertebrates face stressful events, the hypothalamic-pituitary-adrenal (HPA) axis is activated, generating a rapid increase in circulating glucocorticoid (GC) stress hormones followed by a return to baseline levels. However, repeated activation of HPA axis may lead to increase in oxidative stress. One target of oxidative stress is telomeres, nucleoprotein complexes at the end of chromosomes that shorten at each cell division. The susceptibility of telomeres to oxidizing molecules has led to the hypothesis that increased GC levels boost telomere shortening, but studies on this link are scanty. We studied if, in barn swallows Hirundo rustica, changes in adult erythrocyte telomere length between 2 consecutive breeding seasons are related to corticosterone (CORT) (the main avian GC) stress response induced by a standard capture-restraint protocol.
Within-individual telomere length did not significantly change between consecutive breeding seasons. Second-year individuals showed the highest increase in circulating CORT concentrations following restraint. Moreover, we found a decline in female stress response along the breeding season. In addition, telomere shortening covaried with the stress response: a delayed activation of the negative feedback loop terminating the stress response was associated with greater telomere attrition. Hence, among-individual variation in stress response may affect telomere dynamics.

Context dependent variation in corticosterone and phenotypic divergence of Rana arvalis populations along an acidification gradient

Background: Physiological processes, as immediate responses to the environment, are important mechanisms of phenotypic plasticity and can influence evolution at ecological time scales. In stressful environments, physiological stress responses of individuals are initiated and integrated via the release of hormones, such as corticosterone (CORT). In vertebrates, CORT influences energy metabolism and resource allocation to multiple fitness traits (e.g. growth and morphology) and can be an important mediator of rapid adaptation to environmental stress, such as acidification. The moor frog, Rana arvalis, shows adaptive divergence in larval life-histories and predator defense traits along an acidification gradient in Sweden. Here we take a first step to understanding the role of CORT in this adaptive divergence. We conducted a fully factorial laboratory experiment and reared tadpoles from three populations (one acidic, one neutral and one intermediate pH origin) in two pH treatments (Acid versus Neutral pH) from hatching to metamorphosis. We tested how the populations differ in tadpole CORT profiles and how CORT is associated with tadpole life-history and morphological traits.
Results: We found clear differences among the populations in CORT profiles across different developmental stages, but only weak effects of pH treatment on CORT. Tadpoles from the acid origin population had, on average, lower CORT levels than tadpoles from the neutral origin population, and the intermediate pH origin population had intermediate CORT levels. Overall, tadpoles with higher CORT levels developed faster and had shorter and shallower tails, as well as shallower tail muscles.
Conclusions: Our common garden results indicate among population divergence in CORT levels, likely reflecting acidification mediated divergent selection on tadpole physiology, concomitant to selection on larval life-histories and morphology. However, CORT levels were highly environmental context dependent. Jointly these results indicate a potential role for CORT as a mediator of multi-trait divergence along environmental stress gradients in natural populations. At the same time, the population level differences and high context dependency in CORT levels suggest that snapshot assessment of CORT in nature may not be reliable bioindicators of stress.

RNA-seq based transcriptome analysis of ethanol extract of saffron protective effect against corticosterone-induced PC12 cell injury

Background: At present, oral antidepressants are commonly used in the clinical treatment of depression. However, the current drug treatment may lead to more serious adverse reactions. Therefore, we focus on Chinese traditional medicine, trying to find an effective and safe alternative or complementary medicine. Crocus sativus (saffron) is a traditional Chinese herbal medicine, which is typically used in the clinic to regulate anxiety, insomnia, amnesia, and other mental disorder. The study aimed to explore the neuroprotective effect of ethanol extract of saffron (EES) on corticosterone (CORT)- induced injury in PC12 cells and further explored its potential mechanism.
Methods: The authenticity of saffron and the active components of EES were identified by a water test and ultra-performance liquid chromatography-time of flight mass spectrometry system. The screening of cytotoxicity for PC12 cells was incubated with EES in different concentrations for 24 h, and the protective efficacy of EES on CORT (500 μM) -induced PC12 cell injury, cell viability was assessed by Cell Counting Kit-8 (CCK-8) assay. The differentially expressed genes (DEGs) of EES-protected PC12 cells were analyzed using the RNA-seq method, and the results were analyzed for GO and KEGG enrichment. The results of RNA-seq were verified by qPCR analysis.
Results: The saffron was initially identified as authentic in the water test and 10 compounds were identified by Ultra Performance Liquid Chromatography (UPLC)- Mass Spectrometry (MS). The results of CCK-8 demonstrated that EES at concentrations above 640 μg/mL exerted a certain cytotoxic effect, and PC12 cells pretreated with EES (20, 40, and 80 μg/mL) significantly reversed the 500 μM CORT-induced cell death. RNA-seq analysis showed that EES regulated 246 differential genes, which were mainly enriched in the MAPK signaling pathway. Dusp5, Dusp6, Gadd45b, Gadd45G, and Pdgfc were further validated by qPCR. Experimental data showed that the results of qPCR were consistent with RNA-seq.
Conclusions: These findings provide an innovative understanding of the molecular mechanism of the protective effect of EES on PC12 cells at the molecular transcription level, and Dusp5, Dusp6, Gadd45b, Gadd45g, and Pdgfc may be potential novel targets for antidepressant treatment.

Porcine corticosterone / corticosterone (CORT) ELISA Kit
QY-E40120 Qayee Biotechnology 96T 480 EUR

Corticosterone
540089 MedKoo Biosciences 500.0mg 190 EUR

Corticosterone
B7469-5.1 ApexBio 10 mM (in 1mL DMSO) 40 EUR

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Corticosterone and Adrenocorticotrophic Hormone Secretion Is Recovered after Immune Challenge or Acute Restraint Stress in Sepsis Survivor Animals

Background: Clinical and experimental studies report a dysregulation of hypothalamus-pituitary-adrenal (HPA) axis during sepsis that causes impairment in hormone secretion in the late phase contributing for the pathophysiology of the disease. However, it is unclear whether this alteration persists even after the disease remission.
Methods: We evaluated the effect of an immune challenge or restraint stress on the hormone secretion of HPA axis in sepsis survivor rats. Sepsis was induced by cecal ligation-puncture (CLP) surgery. Naive or animals that survive 5 or 10 days after CLP were submitted to lipopolysaccharide (LPS) injection or restraint stress. After 60 min, blood was collected for plasma nitrate, cytokines, adrenocorticotropic hormone (ACTH), and corticosterone (CORT) and brain for synaptophysin and hypothalamic cytokines.
Results: Five days survivor animals showed increased plasma nitrate (p < 0.001) and interleukin (IL)-1β levels (p < 0.05) that were abolished in the 10 days survivors. In the hypothalamus of both survivors, the reverse was seen with IL-6 increased (p < 0.01), while IL-1β did not show any alteration. Synaptophysin expression was reduced in both survivors and did not change after any stimuli. Only the LPS administration increased plasma and/or inflammatory mediators levels in both groups (survivors and naive) being apparently lower in the survivors. There was no difference in the increased secretion pattern of ACTH and CORT observed in the naive and sepsis survivor animals submitted to immune challenge or restraint stress.
Conclusion: We conclude that the HPA axis is already recovered soon after 5 days of sepsis induction responding with normal secretion of ACTH and CORT when required.