The aim of this study was to analyze the expression profiles of the microRNAs (miRNAs) miR-145, miR-181c, miR-199a and miR-1183 in the hippocampus and blood of patients with mesial temporal lobe ...epilepsy with hippocampal sclerosis (MTLE-HS) and to investigate whether these can be used as diagnosis and prognosis biomarkers for epilepsy. Hippocampus and blood samples were collected from 20 patients with MTLE-HS, ten of whom had a favorable surgical outcome (Engel I) and ten with an unfavorable surgical outcome (Engel III-IV). Hippocampus samples from autopsied individuals with no neurological or psychiatric medical history (necropsy samples) and blood samples from healthy individuals were used as controls. Real-time quantitative PCR (RQ-PCR) was used to analyze miRNA expression. The results showed that the expressions of these miRNAs differed quantitatively in the hippocampus and blood of patients with MTLE-HS in comparison to the respective control. This difference was most pronounced for miR-145, which was hypo-expressed in the hippocampus and hyper-expressed in the blood of MTLE-HS patients. MiRNAs miR-145, miR-181c, miR-199a and miR-1183 were hyper-expressed in the blood of patients with MTLE-HS. No statistical differences in the levels of these miRNAs in the blood or hippocampus were found between Engel I patients and Engel III-IV patients. These results suggest that the analyzed microRNAs are potential circulating biomarkers for epilepsy diagnosis.
•Literature highlights the role of miR-27a-3p, miR-328-3p and miR-654-3p in epilepsy.•MiR-328-3p is the most important peripheral biomarker for the diagnose of MTLE-HS.•Circulating miR-654-3p is a ...biomarker for epilepsy surgery prognosis.
MicroRNAs have been progressively investigated as post-transcriptional regulators playing important roles in epilepsy pathophysiology. Here we investigate three promising microRNAs (miR-27a-3p, miR-328-3p and miR-654-3p) previously described in the literature as possible peripheral biomarkers for epilepsy diagnose and surgical prognosis. Serum samples from 28 patients with mesial temporal lobe epilepsy with hippocampal sclerosis (MTLE-HS) were analyzed, 14 with good surgical prognosis (Engel I) and 14 with unfavorable surgical prognosis (Engel III-IV). Serum samples from 11 healthy volunteers were the control group. The microRNAs expression analysis was performed using real-time PCR. The present results did not endorse the role of miR-27a-3p as a peripheral biomarker for epilepsy diagnosis or surgical prognosis. MiR-328-3p, however, presented significant area under the curve (AUC) values when comparing controls to Engel I (90.3%), controls to Engel III-IV (96.8%) and controls to Engel I + Engel III-IV (i.e., epilepsy patients, AUC = 93.5%). Additionally, miR-654-3p displayed AUC = 74.7% when comparing controls to Engel I patients (p = 0.004), and AUC = 73.6% (p = 0.04) in the attempt to discriminate unfavorable from favorable surgical prognosis. In conclusion, the ANOVA and ROC analyzes with the respective AUC, specificity and sensitivity values allows us to conclude that miR-328-3p is the most important peripheral biomarker for the diagnosis of MTLE-HS. In terms of predicting the surgical prognosis of MTLE-HS patients, miR-654-3p proved to be the only microRNA evaluated to present statistical power to differentiate, as a peripheral biomarker, Engel I from Engel III-IV patients.
This study aimed to analyze the cerebellum of rats submitted to an experimental focal cerebral ischemia, by middle cerebral artery occlusion for 90 minutes, followed by reperfusion for 48 hours, ...associated with an alcoholism model.
Fifty adult Wistar rats were used, subdivided into five experimental groups: control group (C): animals submitted to anesthesia only; sham group (S): animals submitted to complete simulation of the surgical procedure; ischemic group (I): animals submitted to focal cerebral ischemia for 90 minutes followed by reperfusion for 48 hours; alcoholic group (A): animals that received daily absolute ethanol diluted 20% in water for four weeks; and, ischemic and alcoholic group (I + A): animals receiving the same treatment as group A and, after four weeks, submitted to focal cerebral ischemia for 90 minutes, followed by reperfusion for 48 hours. The cerebellum samples were collected and immunohistochemical analysis of Caspase-9 protein and serum analysis by RT-PCR of microRNAs miR-21, miR-126 and miR155 were performed.
The expression of Caspase-9 was higher in groups I, A and I + A. In the microRNAs analyses, miR-126 was higher in groups A and I + A, miR-155 was higher in groups I and I + A.
We conclude that apoptosis occurs in the cerebellar cortex, even if it is distant from the ischemic focus, and that microRNAs 126 and 155 show a correlation with cellular apoptosis in ischemic rats and those submitted to the chronic alcohol model.
Abstract Purpose To evaluate the effects of alcohol exposure and diabetes on apoptotic process in the corpus cavernosum. Methods Forty eight male Wistar rats were divided into four groups: control, ...diabetic, alcoholic and diabetic-alcoholic. Samples of the corpus cavernosum were prepared to study protein expression of apoptotic genes (Caspases-3 and 9) by immunohistochemistry and Real-Time PCR. Results The immunoreactivity of Caspases-3 and -9 was diffuse and higher in the treated groups though there was no significant difference between the experimental groups, only when compared with the control group. An increase was observed in the gene expression of Caspases-9 in the diabetic and ethanol-diabetic groups when compared with control and ethanol groups. Conclusions The association of these factors (ethanol and diabetes) probably can affect the apoptosis mechanism in lesions of the cavernous tissue in the rat penis. Both gene and protein expression of Caspase-9 in diabetic and ethanol-diabetic groups suggest the involvement of the apoptosis cascade from this study model.
Abstract Purpose To evaluate the effect of chronic alcoholism on morphometry and apoptosis mechanism and correlate with miRNA-21 expression in the corpus cavernosum of rats. Methods Twenty-four rats ...were divided into two experimental groups: Control (C) and Alcoholic group (A). After two weeks of an adaptive phase, rats from group A received only ethanol solution (20%) during 7 weeks. The morphometric and caspase-3 immunohistochemistry analysis were performed in the corpus cavernosum. The miRNA-21 expression was analyzed in blood and cavernous tissue. Results Chronic ethanol consumption decreased cavernosal smooth muscle area of alcoholic rats. The protein expression of caspase 3 in the corpus cavernosum was higher in A compared to the C group. There was no difference in the expression of miRNA-21 in serum and cavernous tissue between the groups. Conclusion Chronic ethanol consumption reduced smooth muscle area and increased caspase 3 in the corpus cavernosum of rats, without altered serum and cavernosal miR-21 gene expression.
Diagnosis and treatment of pancreatic ductal adenocarcinoma (PA) remains a challenge in clinical practice. The aim of this study was to assess the role of microRNAs (miRNAs-21, -23a, -100, -107, ...-181c, -210) in plasma and tissue as possible biomarkers in the diagnosis of PA. Samples of plasma (PAp-n = 13), pancreatic tumors (PAt-n = 18), peritumoral regions (PPT-n = 9) were collected from patients during the surgical procedure. The control group consisted of samples from patients submitted to pancreatic surgery for trauma or cadaveric organs (PC-n = 7) and healthy volunteers donated blood (PCp-n = 6). The expression profile of microRNAs was measured in all groups using RT-PCR, serum CA19-9 levels were determined in PA and PC. In tissue samples, there was a difference in the expression of miRNAs-21, -210 (
p
< 0.05) across the PAt, PC and PPT groups. The PAp showed overexpression of miRNAs-181c, -210 (
p
< 0.05) when compared to PCp. The combination of miRNAs-21, -210 tissue expression and serum CA19-9 showed 100% accuracy in the diagnosis of PA, as well as miR-181c expression in the plasma (PApxPCp). The expression of microRNAs in plasma proved to be a promising tool for a noninvasive detection test for PA, as well as further studies will evaluate the utility of microRNAs expression as biomarkers for prognostic and response to therapy in PA.
•MiR-219 and NMDA-NR1 expressions are inverse both for amygdala and hippocampus.•NR1 and GluR2 were upregulated in the amygdala of patients with epilepsy.•MiR-219 may play a regulatory role in ...excitatory neurotransmission in epilepsy.
Mesial temporal lobe epilepsy with hippocampal sclerosis is the most frequent form of focal epilepsy in adults, and it is often refractory to drug treatment. Regardless of the efforts on developing new antiepileptic drugs for refractory cases, studies suggest a need for better understanding the molecular bases of epilepsy. The microRNAs have been progressively investigated as potential targets for both epilepsy mechanisms elucidation and treatment. Therefore, the goal of this study was to evaluate the differential expression of miR-219, miR-181b, and miR-195, previously described as regulators of the excitatory neurotransmitter receptors NMDA-R1 and AMPA-GluR2 and inhibitory neurotransmitter GABAA (α2, β3, and γ2 subunits) in the amygdala and hippocampus of patients with mesial temporal lobe epilepsy. Based on genes and miRNAs’ quantitative Polymerase Chain Reaction (qPCR) from 18 patients with epilepsy, our results showed an inverse relationship between miR-219 and NMDA-NR1 expression in both the amygdala and hippocampus in comparison to their expression in controls. NR1 and GluR2 were upregulated in the amygdala of epileptic patients. Low miR-195 expression was observed in the amygdala of patients with epilepsy. Our findings indicate that miR-219 has a possible regulatory role in excitatory neurotransmission in patients with epilepsy, contributing to the new avenue of miRNA biology in drug-resistant epilepsy, reserving huge potential for future applications and clinical interventions in conjunction with existing therapies.
Introduction
Pancreatic adenocarcinoma (PA) remains one of the most aggressive malignant tumors of the digestive tract, with a five‐year survival rate of 5%. Although early detection offers the best ...chance to cure pancreatic adenocarcinoma, early stage disease presents little or no symptoms. The most commonly used marker for early detection of AP is serum CA19‐9, which has a sensitivity of 70% to 80%, however, with a specificity less than 50%. MiRNAs, endogenous small RNAs of approximately 17–25 nucleotides, present an altered expression in various cancers, suggesting its close relationship to disease processes, with the possibility of being a molecular biomarker for diagnosis and prognosis of cancer.
Objectives
To evaluate the effect of the combination of the association of anti‐apoptotic microRNAs: miR‐15a and miR‐16 and pro‐apoptotic microRNAs: miR‐21, miR‐221 and miR‐222 expression profile and the protein XIAP with serum CA19‐9 in diagnosis of diagnosis of pancreatic adenocarcinoma.
Patients and Methods
We studied 23 samples of PA and 20 as control samples: 13 of normal pancreatic tissue adjacent to the tumor (PPT), collected during pancreatic resections and 7 normal pancreas (NP) from deceased donors or patients who have undergone pancreatectomy affected by trauma. CA19‐9 was measured in serum and checked for expression of the miRNAs and XIAP in PA, PPT and NP through amplification by real‐time quantitative PCR (RT‐PCR). The usefulness of miRNAs and XIAP in the diagnosis of PA, alone or associated with CA19‐9, were analyzed by the construction of ROC curves.
Results
It was observed significant differences in the expression of miR‐21 in the comparison between the PA and NP groups (p<0.05). However, there was no significant difference in the expression of the protein XIAP between groups, as well as the expression of miR‐21 between groups NP and PPT. The combination of miR‐21 with CA19‐9 was more effective to discriminate the PA and NP (AUC‐ROC: 0.94; sensitivity: 78%, specificity: 99%) when compared with the assessments isolated.
Conclusion
The expression of miR‐21 was demonstrated to be effective in discriminating between PA and NP and the association between miR‐21 and CA19‐9 increased the sensitivity and accuracy in the diagnosis of PA in the sample studied.
This is from the Experimental Biology 2019 Meeting. There is no full text article associated with this published in The FASEB Journal.
BackgroundGlioblastoma is an incurable neoplasm. Its hypoxia mechanism associated with cancer stem cells (CSCs) demonstrates hypoxia-inducible factor 1α (HIF-1α) expression regulation, which is ...directly related to tumor malignancy. The aim of this study was to identify a possible tumor malignancy signature associated with regulation of HIF-1α by microRNAs miR-21 and miR-326 in the subpopulation of tumor stem cells which were irradiated by ion in primary culture of patients diagnosed with glioblastoma. Materials and methodsWe used cellular cultures from surgery biopsies of ten patients with glioblastoma. MicroRNA expressions were analyzed through real-time polymerase chain reaction (PCR ) and correlated with mortality and recurrence. The ROC curve displayed the cutoff point of the respective microRNAs in relation to the clinical prognosis, separating them by group. ResultsThe miR-21 addressed high level of expression in the irradiated neurosphere group (p = 0.0028). However, miR-21 was not associated with recurrence and mortality. miR-326 can be associated with tumoral recurrence (p = 0.032) in both groups; every 0.5 units of miR-326 increased the chances of recurrence by 1,024 (2.4%). ConclusionThe high expression of miR-21 in the irradiated group suggests its role in the regulation of HIF-1α and in the radioresistant neurospheres. miR-326 increased the chances of recurrence in both groups, also demonstrating that positive regulation from miR-326 does not depend on ionizing radiation treatment.
Introduction
Ethanol consumption has been described to increase the levels of plasma endothelin‐1 (ET‐1) suggesting that chronic consumption activates endothelinergic pathway and also to affect ...nitric oxide synthase (NOS) derived nitric oxide (NO) and NOS expression in different tissues. Recent evidence showed that chronic ethanol consumption increases ET‐1‐induced sustained contraction of trabecular smooth muscles cells of the corpora cavernosal in corpus cavernosum of rat by mechanism that involves increased expression of ETA eee and ETB receptors.
Objectives
To evaluate the effects of chronic ethanol consumption associated with diabetes affected the endothelin system in the cavernosal smooth muscle (CSM) of rats, as well as the effects of chronic ethanol consumption and diabetes on nitric oxide (NO)‐mediated relaxation of cavernosal smooth muscle (CSM).
Materials and Methods
Corpus cavernosum were divided into four groups: control, ethanol, diabetic and ethanol‐diabetic of Male Wistar rats. We analyzed protein and gene expression of endothelial NO synthase (eNOS), inducible NO synthase (iNOS) and endothelin receptors (ETA eeeand ETB) by immunohistochemistry and real time‐PCR. The expression of miRNAs‐27a, 199 and 155 by real time‐PCR in corpus cavernosum. It was also evaluated the effect of ethanol consumption associated with diabetes on the relaxation induced by acetylcholine (ACh; 0.01–1000 micromol/L) and in the contraction induced by Endothelin‐1 (ET‐1 0.1 nmol‐ 1μmol L‐1) in organ chambers for measurement of isometric tension.
Results
The endothelium‐dependent relaxation induced by ACh was decreased in CSM from ethanol, diabetic, ethanol‐diabetic groups when compared with the control group. Immunohistochemistry for eNOS, iNOS, ETA eeeand ETB showed an increase in cavernosal smooth muscle cells in the ethanol, diabetic and ethanol‐diabetic groups when compared with the control group. Similarly, the mRNA levels for eNOs, ETA eeeand ETB were increased in CSM of groups ethanol, diabetic and ethanol‐diabetic and miRNAs‐27a, 155 and 199 were decreased in cavernosal smooth muscle from the ethanol, diabetic and ethanol‐diabetic groups.
Conclusions
Our results showed that chronic ethanol consumption associated with diabetes was an impairment of relaxation of CSM from ethanol, diabetic, and ethanol‐diabetic rats that involved a decrease in the NO pathway by endothelium‐dependent mechanisms accompanied by a change in the corpus cavernosum contractile sensitivity. We can also suggest that chronic ethanol consumption associated with diabetes changed the endothelinergic system in the CSM played a role in the pathogenesis of erectile dysfunction, suggesting that these changes of endothelin receptors were regulated by miRNAs‐155, 199.
This is from the Experimental Biology 2018 Meeting. There is no full text article associated with this published in The FASEB Journal.
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