Volatile Organic Compounds in Patients With Acute Kidney Injury and Changes During Dialysis

Objectives: To characterize volatile organic compounds in breath exhaled by ventilated care patients with acute kidney injury and changes over time during dialysis. Design: Prospective observational feasibility study. Setting: Critically ill patients on an ICU in a University Hospital, Germany. Patients: Twenty sedated, intubated, and mechanically ventilated patients with acute kidney injury and indication for dialysis. Interventions: Patients exhalome was evaluated from at least 30 minutes before to 7 hours after beginning of continuous venovenous hemodialysis. Measurements and Main Results: Expired air samples were aspirated from the breathing circuit at 20-minute intervals and analyzed using multicapillary column ion-mobility spectrometry. Volatile organic compound intensities were compared with a ventilated control group with normal renal function. A total of 60 different signals were detected by multicapillary column ion-mobility spectrometry, of which 44 could be identified. Thirty-four volatiles decreased during hemodialysis, whereas 26 remained unaffected. Forty-five signals showed significant higher intensities in patients with acute kidney injury compared with control patients with normal renal function. Among these, 30 decreased significantly during hemodialysis. Volatile cyclohexanol (23 mV; 25–75th, 19–38), 3-hydroxy-2-butanone (16 mV, 9–26), 3-methylbutanal (20 mV; 14–26), and dimer of isoprene (26 mV; 18–32) showed significant higher intensities in acute kidney impairment compared with control group (12 mV; 10–16 and 8 mV; 7–14 and not detectable and 4 mV; 0–6; p

https://ift.tt/2EGzn0C

The function of hyperpolarization-activated cyclic nucleotide-gated channel in diabetic cystopathy

OBJECTIVE: We aimed at investigating changes in the expression and physiological function of hyperpolarization-activated cyclic nucleotide-gated (HCN) channels in interstitial cells of Cajal (ICC) in diabetic state.

MATERIALS AND METHODS: Twenty adult female Sprague-Dawley (SD) rats were randomly assigned to control and Zucker diabetic fatty (ZDF) group. The protein and mRNA expression of HCN isoforms and C-kit in the rat bladders were detected using Western blotting and reverse transcription-polymerase chain reaction (RT-PCR). The bladder contraction was evaluated using a bladder smooth muscle strip test. Whole cell patch-clamp techniques were used to detect the activity of HCN channels. Immunofluorescent staining was used to the positive expression of HCN and C-kit in ICC.

RESULTS: cAMP, as HCN channel-specific stimulant, could increase the frequency and amplitude of spontaneous contractions in both group, while cAMP inducing contraction of ZDF rats, was still significantly lower compared with the control group. Acute bladder ICCs were isolated by collagenase digestion. Classic Ih current pattern was recorded on ICCs while Ih current amplitude of ICCs from ZDF diabetic rats was significantly lower than the control group. The expression and mRNA of HCN1-4 isoforms in ZDF diabetic rats were both significantly lower compared with the control group. Meanwhile, the number of c-kit positive cells in ZDF diabetic rats showed no significant differences compared with controls. The morphological structure of ICC in the bladder of ZDF rats was relatively loose and the number of their cell process was apparently decreased.

CONCLUSIONS: The structure of ICCs in ZDF rats was relatively loose, their connection to each other was also diminished. The expression of HCN was down-regulated and its response to cAMP was also decreased. HCN channels in bladder ICCs might regulate detrusor contraction. Changes in HCN expression and activity in bladder ICCs might be one of the most important mechanisms of diabetic cystopathy.

L'articolo The function of hyperpolarization-activated cyclic nucleotide-gated channel in diabetic cystopathy sembra essere il primo su European Review.

https://ift.tt/2yzsvwy

Comment on “Neuronal nitric oxide synthase inhibition reduces brain damage by promoting collateral recruitment” in a cerebral hypoxia-ischemia mice model

L'articolo Comment on "Neuronal nitric oxide synthase inhibition reduces brain damage by promoting collateral recruitment" in a cerebral hypoxia-ischemia mice model sembra essere il primo su European Review.

https://ift.tt/2O9Pkfv

The involvement of miR-155 in blood pressure regulation in pregnant hypertension rat via targeting FOXO3a

OBJECTIVE: Pathogenesis factor of pregnant hypertension is still unclear and lacks of effective treatment. MiR-155 is a recently discovered miRNA molecule with differential expression in pregnant hypertension, which participates in the disease regulation. As a downstream target gene of miR-155, FOXO3a is correlated with blood pressure regulation. We investigated the regulatory role and mechanism of miR-155 in pregnant hypertension.

MATERIALS AND METHODS: We established a pregnant hypertension rat model, on which miR-155 inhibitor or FOXO3a siRNA was applied, followed by HE staining, 24 h urea protein, blood pressure and serum creatine assay to evaluate disease severity.

RESULTS: MiR-155 expression was significantly elevated in model rats, accompanied by a reduction of the FOXO3a level. MiR-155 inhibitor suppressed miR-155 expression, increased FOXO3a level and placental tissue morphology by HE staining, and depressed blood pressure as well as serum creatine level. Downregulation of FOXO3a by specific siRNA resulted in opposite effects. These results illustrated the miR-155 mediated FOXO3a expression in pregnant hypertension.

CONCLUSIONS: The inhibition of miR-155 improves the damage of pregnant hypertension via the upregulation of FOXO3a, which provides academic leads for the future therapy of pregnant hypertension.

L'articolo The involvement of miR-155 in blood pressure regulation in pregnant hypertension rat via targeting FOXO3a sembra essere il primo su European Review.

https://ift.tt/2O9P8Nj

Up-regulation of miR-517-5p inhibits ERK/MMP-2 pathway: potential role in preeclampsia

OBJECTIVE: To investigate the potential role of miRNA-517-5p in preeclampsia and its underlying mechanism.

MATERIALS AND METHODS: Placenta samples were obtained from 20 women with preeclampsia and 20 women with normal pregnancies. Expression level of miR-517-5p in placenta samples and JAR cells was detected. MiRNA-517-5p mimics or inhibitor was transfected in JAR cells, followed by detection of proliferative and invasive abilities of JAR cells. In addition, the expressions of extracellular regulated protein kinases (ERK), phospho-extracellular regulated protein kinases (p-ERK) and matrix metalloproteinase-2 (MMP-2) in JAR cells were evaluated by Western blot. Meanwhile, the mRNA level of MMP-2 was evaluated by Real-time polymerase chain reaction (PCR). The luciferase assay was applied to identify the target gene of miRNA-517-5p.

RESULTS: Increased level of miR-517-5p was detected in placenta samples of preeclampsia patients compared with normal pregnancies. MiRNA-517-5p could regulate proliferative and invasive abilities of JAR cells. Furthermore, miRNA-517-5p could regulate ERK/MMP-2 pathway in JAR cells, which would contribute to the pathophysiology of preeclampsia. The luciferase assay showed MMP-2 was the target gene of miR-517-5p. Further studies showed that MMP-2 was dysregulated in preeclampsia.

CONCLUSIONS: MiR-517-5p is highly expressed in placenta samples of preeclampsia pregnancies, which could promote proliferative and invasive abilities of JAR cells by inhibiting ERK/MMP-2 pathway.

L'articolo Up-regulation of miR-517-5p inhibits ERK/MMP-2 pathway: potential role in preeclampsia sembra essere il primo su European Review.

https://ift.tt/2yDIFoC

LncRNA HOTAIR participates in the development and progression of adrenocortical carcinoma via regulating cell cycle

OBJECTIVE: To explore the role of HOTAIR in the pathogenesis of adrenocortical carcinoma (ACC) and its underlying mechanism.

PATIENTS AND METHODS: Differentially expressed lncRNA (HOTAIR) in ACC was screened out from the GEO database. The survival analysis and ROC curve were performed according to HOTAIR expressions in ACC patients. The correlation between HOTAIR expression and clinical information of ACC patients was analyzed by chi-square test. The univariate and multivariate COX regression analysis was carried out to analyze the relationship between HOTAIR expression, disease-free survival (DFS) and overall survival (OS) of ACC patients. We then detected HOTAIR expression in 77 ACC tissues and 30 normal tissues by qRT-PCR (quantitative Real-time polymerase chain reaction). ACC cell lines were further screened out for the following in vitro experiments. After altering HOTAIR expression in ACC cells by plasmid transfection, proliferation and cell cycle were detected by Cell Counting Kit-8 (CCK-8) and colony formation assay, respectively. Finally, Western blot was utilized to detect expressions of cell cycle-related genes in ACC cells.

RESULTS: HOTAIR was overexpressed in ACC tissues than that of normal tissues. HOTAIR expression was remarkably increased in ACC with T3 and T4 stage than that of T1 and T2 stage. Moreover, HOTAIR expression was remarkably increased in ACC with stage III and IV than that of stage I and II. HOTAIR was an independent prognostic factor for DFS and OS of ACC patients. For in vitro experiments, inhibited proliferation and arrested cell cycle were observed in H295R cells transfected with si-HOTAIR. Opposite results were obtained after SW-13 cells were transfected with HOTAIR overexpression plasmid. Furthermore, expressions of cell cycle-related genes, including Cyclin D1, p-Rb and p-GSK3β were remarkably decreased after HOTAIR knockdown.

CONCLUSIONS: We demonstrated for the first time that HOTAIR is overexpressed in ACC and is a prognostic risk factor in ACC patients. HOTAIR participates in the development and progression of ACC via shortening cell cycle and promoting proliferation of ACC cells.

L'articolo LncRNA HOTAIR participates in the development and progression of adrenocortical carcinoma via regulating cell cycle sembra essere il primo su European Review.

https://ift.tt/2O9P2oV

Effect of hypoxia on the Twist1 in EMT of cervical cancer cells

OBJECTIVE: To explore the effect of hypoxia on the Twist1 expression in epithelial-mesenchymal transition of the cervical cancer cells.

MATERIALS AND METHODS: In this study, we simulated the normoxia and hypoxia environment, where HeLa cells were cultured, respectively. Cell invasion ability was measured by the transwell assay, while the GLI-1 protein and mRNA expressions were measured by Real-time polymerase chain reaction (RT-PCR) and Western blot assays. After that, HeLa cells were transfected with the GLI-1-specific siRNA, followed by the measurement of mRNA and protein expressions using RT-PCR and Western blot assays, as well as the cell invasion ability by the transwell assay.

RESULTS: We found that in hypoxic environment, GLI-1 was up-regulated in HeLa cells, with enhanced invasion ability. However, silencing the expression of GLI-1 could reverse the up-regulation of GLI-1 compromising the invasion ability of HeLa cells.

CONCLUSIONS: Hypoxia may account for the increased invasion of HeLa cells, which is realized by the up-regulated GLI-1.

L'articolo Effect of hypoxia on the Twist1 in EMT of cervical cancer cells sembra essere il primo su European Review.

https://ift.tt/2yztkWk