This study aimed to investigate the morphometric and the pattern of protein and gene expression related to the extrinsic apoptotic pathway in experimental focal cerebral ischemia and the hole of ...neuroprotection with hypothermia and ketoprofen. For this analysis, 120 rats were randomly divided into 3 groups (20 animals each): control - no surgery (20 animals); sham - simulation of surgery (20 animals); ischemic - focal ischemia for 1 hour, without reperfusion (80 animals) and divided into four subgroups with 20 animals each: ischemic + intraischemic hypothermia; ischemic + previous intravenous ketoprofen, and ischemic + hypothermia and ketoprofen. The infarct volume was measured using morphometric analysis of infarct areas defined by triphenyl tetrazolium chloride and the patterns of expression of the apoptosis genes (Fas, c-Flip, caspase-8 and caspase-3) and the apoptosis protein caspase-3 were evaluated by quantitative real-time PCR and immunohistochemistry, respectively. Hypo expression of genes of extrinsic pathway of apoptosis was observed: Fas receptor, c-Flip and caspase-8 in the ischemics areas. Increases in the gene and protein caspase-3 in the ischemic areas were also observed, and these increases were reduced by hypothermia and ketoprofen, also noted in the morphometric study. The caspases-3 increase suggests that this gene plays an important role in apoptosis, probably culminating in cell death and that the neuroprotective effect of hypothermia and ketoprofen is involved.
The chronic consumption of alcohol causes a worsening of the events that follow the cerebral ischemia. These events are regulated through the expression of several genes and microRNAs. The aimof this ...work was To analyze and describe the expression profile of PARP and AIF and miRNA-9 proteins in rats submitted to focal cerebral ischemia, associated or not with chronic alcoholism model. Methods: Twenty adult Wistar rats, subdivided into: control; ischemic; alcoholic and ischemic / alcoholized for immunohistochemical analysis and miRNA-9 gene expression. Results: There was a reduction in the protein expression of PARP-1 and a positive marking for AIF in the ischemic / alcoholized group. The miRNA-9 did not obtain significant expression. The association of ischemia with chronic alcohol use promoted a tendency to low expression of miRNA-9, low expression of PARP-1 and high expression of AIF, indicating an interference in the protective effect of miRNA-9 be observed in the other groups.
•miR-143 is related with more inflammation and edema of intestinal loops of gastroschisis rat model.•Time of exposure of the amniotic fluid increases the intestinal damage.•miR-143 is a possible ...marker of gut inflammation.
Gastroschisis (GS) is a congenital anomaly in the abdominal wall with the intestinal loops exiting laterally to the umbilicus. The contact of the loops with Amniotic Fluid (AF) causes an inflammatory process in the exposed part, leading to an extended hospital stay and an increased risk of morbidity due to alterations related to intestinal motility. The authors aimed to evaluate the time of exposure to the AF in the experimental GS and to search for potential biomarkers of intestinal inflammation by measuring microRNAs.
Rat fetuses were divided into three groups: a) CONTROL, b) GS reared on day 18 (GS = 18), and c) GS reared on day 19.5 (GS = 19) (term = 22 days). On day 21.5, the fetuses were removed for biometric parameters and biochemical analyses: 1) Biometrics: Body and Intestinal Weight (BW, IW), and intestinal-body weight ratio (IW/BW); 2) Descriptive histopathology and 3) miR-143 quantification by real-time Polymerase Chain Reaction (PCR).
BW was higher in CONTROL than GS 18 and G19 (p < 0.05). IW, IW/BW, intestinal water, and mRNA-143 were higher in GS 18 and GS 19 than in CONTROL, and GS 18 was higher than GS 19 (p < 0.05). The average of the inflammation score from the intestinal wall with mucosal inflammation and intra-epithelial lymphocytes shows worst in GS 18 and GS 19 vs. CONTROL (p < 0.05).
The tissue expression of mRNA-143 and the morphological changes in the intestine of GS worsened according to the time of exposure to AF, which could be a possible marker of fetal intestinal damage.
•miR-629-3p, miR-1202 and miR-1225-5p are involved in GABAergic pathways.•miR-629-3p, miR-1202 and miR-1225-5p are hyper-expressed in epilepsy patients’ blood.•miR-629-3p, miR-1202 and miR-1225-5p ...are potential diagnostic biomarkers of MTLE-HS.•miR-1202 may be a potential surgery prognose biomarker in amygdala of MTLE-HS cases.
Mesial temporal lobe epilepsy (MTLE) is a symptomatic epilepsy syndrome clinically characterized by high prevalence, pharmacoresistance, good surgical prognosis and hippocampal sclerosis (HS); however, no singular criteria can be considered sufficient for the MTLE-HS diagnosis. MicroRNAs (miRNAs) are small non-coding molecules that act as important gene-expression regulators at post-transcriptional level. Evidences on the involvement of miRNAs in epilepsy pathogenesis as well as their potential to be employed as biomarkers claim for investigations on miRNAs’ applicability as epilepsy diagnosis and prognosis biomarkers. Consequently, the present study aimed to evaluate the applicability of three specific miRNAs as biomarkers of diagnosis and surgical outcomes in adult patients with MTLE-HS.
Hippocampus, amygdala and blood samples from 20 patients with MTLE-HS were analyzed, 10 with favorable surgical prognosis (Engel I) and 10 with unfavorable surgical prognosis (Engel III-IV). For the control groups, hippocampus and amygdala from necropsy and blood samples from healthy individuals were adopted. The miRNAs expression analysis was performed using Real-Time Quantitative Polymerase Chain Reaction for miRNAs highlighted from microarray as being involved in GABAergic neurotransmission.
The miRNAs miR-629-3p, miR-1202 and miR-1225-5p were found to be hyper-expressed in MTLE-HS patients’ blood.
Our data suggest the existence of three circulating miRNAs (miR-629-3p, miR-1202 and miR-1225-5p) that could possibly act as additional tools in the set of factors that contribute to MTLE-HS diagnose.
Previous studies from our group described the consequences of using ethanol on penile erection. Nevertheless, the molecular mechanisms surrounding microRNAs, apoptosis process and their relationship ...with erectile dysfunction associated with alcohol consumption are still poorly understood. The objective of this analysis was to evaluate the mechanism of apoptosis by the expression of AIF and PARP, as well as their regulatory microRNAs: miR-145, miR-210 and miR-486, in the corpus cavernosum of rats submitted to a semivoluntary alcoholism model. For this study 24 Wistar rats were divided into two groups: control (C) and treated with 20 % ethanol (A) for seven weeks. The corpus cavernosum samples were prepared for immunohistochemical analysis of AIF and PARP protein expression, and microRNAs miR-145, miR-210, miR-486 gene expression in cavernous tissue was performed by real time PCR. The immunohistochemical analysis showed little nuclear positive labeling for the protein PARP and AIF in the corpus cavernosum of control and ethanol treated animals. After analysis of miR-145, -210 and -486 microRNA expression in the 12 animals studied, no results were found with significant statistical difference between the control and alcoholized groups. The expression of AIF and PARP and their regulatory microRNAs involved in apoptotic process (miR-145, miR-210 and miR-486) were not altered in the corpus cavernosum of rats submitted to semivoluntary alcoholism. KEY WORDS: Alcoholism; Apoptosis; Erectile Dysfunction; MicroRNAs.
Abstract only
Introduction
The differential diagnosis between pancreatic adenocarcinoma (PA) and chronic pancreatitis is still a challenge and the use of biomarkers association can mitigate these ...difficulties.
Objectives
To evaluate the effect of combining miRNA‐21 and serum CA 19‐9 in the diagnosis of PA.
Patients and Methods
We studied 23 PA samples and 20 samples as controls: 13 from normal pancreatic tissue adjacent to the tumor (TPR) collected during pancreatic resections and 7 from normal pancreas (NP) from cadaveric donors or patients undergoing pancreatectomy for trauma. Serum CA 19‐9 was measured and miRNA‐21 expression were verified in PA, NP and TPR by quantitative real‐time polymerase chain reaction (PCR) amplification. The usefulness of miR‐21 in the diagnosis of PA, isolated or associated with CA 19‐9 were analyzed by constructing ROC curves.
Results
There was a significant difference in miR‐21 expression in the comparison between NP and PA groups (p <0.05). However, there was no significant difference in miRNA‐21 expression between NP and TPR groups. The combination of miRNA‐21 with CA 19‐9 was more effective in discriminating PA from NP (AUC‐ROC: = 0.94; sensitivity: 78%; specificity: 99%) when compared with isolated evaluations.
Conclusion
The expression of miRNA‐21 was effective in discriminating between PA and NP and its association with serum CA 19‐9 increased the sensitivity and accuracy in diagnosing PA in the sample studied.
Support or Funding Information
This study was financed in part by the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior ‐ Brasil (CAPES) ‐ Finance Code 001.
•Ionizing radiation and temozolomide reduced the viability of CSCs from GBM patients.•Ionizing radiation and temozolomide were able to modify expression MGMT gene of GCSCs.•miR-181d interferes in the ...CSCs response submitted to standard treatment from GBM.
Despite the increased understanding of the oncological mechanisms underlying Glioblastoma multiforme (GBM) pathophysiology, and recent advances in therapeutic strategies such as maximal surgical resection and post-operative radiotherapy with concomitant and adjuvant temozolomide chemotherapy, the prognosis for patients with brain tumors remains limited. Evidences indicate that the assessment of DNA methylation status in cancer stem cells would allow identifying molecules expressed in these cells, to lead to targeted elimination of this critical population from brain tumors, making the glioblastoma treatment more effective. This study aimed to analyze the role of microRNA-181d associated with the methylation status of the O6-methylguanine methyl transferase (MGMT) gene in Glioblastoma multiforme cancer stem cells subjected to treatment with temozolomide and ionizing radiation. Such responses were analyzed in terms of cell survival, evaluation of the MGMT gene methylation status by MS-HRM (Methylation-Sensitive High Resolution Melting), and analysis of miRNA-181d and MGMT gene expression by relative quantification of mRNA levels in cancer stem cells subjected to treatment with temozolomide and ionizing radiation, isolated or combined. We showed that ionizing radiation and temozolomide reduced the viability of cancer stem cells from GBM patients, as well as modified MGMT gene and miRNA-181d expression in cancer stem cells, suggesting that miRNA-181d interferes in the glioblastoma cancer stem cell response to treatment with temozolomide and ionizing radiation.
Stroke is one of the main causes of death and disability worldwide. The great impact on the quality of life of the population and on the health system justifies that we seek relevant alternatives to ...reduce the incidence and improve the treatment and recovery of patients affected by this disease. Physical exercise appears as an important tool in this scenario, being already pointed out as a possible therapeutic approach for the prevention of non-contagious chronic diseases. In this context, biomarkers such as miRNAs that respond to physical exercise and are directly related to several epigenetic mechanisms appear. Therefore, explaining the molecular mechanisms involved during physical exercise will lead to a better understanding of each stimulus and the dose to be used to better respond to each situation, thus being a promising approach for the evolution of prescription and control of training and processes recovery from various diseases, including stroke. Forty-eight Wistar rats were used, divided into four experimental groups: control group, ischemia group, physical exercise group and exercise + ischemia group. Real-time PCR methodology was used to analyze the expression of miRNAs: miR-126, miR-133b and miR-221. In our study we observed a significant difference in the expression of miR-221 between the control group and the others groups. However, microRNAs: miR-126 and miR-133b do not show significant differences in expression between groups. KEY WORDS: Cerebral ischemia; Physical exercise; MicroRNAs; Apoptosis. El ictus es una de las principales causas de muerte y discapacidad en todo el mundo. El gran impacto en la calidad de vida de la población y en el sistema de salud justifica buscar alternativas pertinentes para reducir la incidencia y mejorar el tratamiento y recuperación de los pacientes afectados por esta enfermedad. El ejercicio físico aparece como una herramienta importante en este escenario, siendo ya señalado como un posible abordaje terapéutico para la prevención de enfermedades crónicas no contagiosas. En este contexto, aparecen biomarcadores como los miRNAs que responden al ejercicio físico y están directamente relacionados con varios mecanismos epigenéticos. Por lo tanto, explicar los mecanismos moleculares involucrados durante el ejercicio físico conducirá a una mejor comprensión de cada estímulo y la dosis a utilizar para responder mejor a cada situación, siendo así un enfoque prometedor para la evolución de la prescripción, el control del entrenamiento y los procesos de recuperación de diversas enfermedades, incluido el accidente cerebrovascular. Se utilizaron cuarenta y ocho ratas Wistar, divididas en cuatro grupos experimentales: grupo control, grupo isquemia, grupo ejercicio físico y grupo ejercicio + isquemia. Se utilizó la metodología de PCR en tiempo real para analizar la expresión de miRNAs: miR-126, miR-133b y miR-221. En nuestro estudio observamos una diferencia significativa en la expresión de miR-221 entre el grupo control y los demás grupos. Sin embargo, los microARN: miR-126 y miR-133b no mostraron diferencias significativas en la expresión entre grupos. PALABRAS CLAVE: Isquemia cerebral; Ejercicio físico; microARN; Apoptosis.
Cerebral ischemia has not only a high mortality rate, which is the second leading cause of death worldwide, but is also responsible for severe disabilities in working age individuals, generating ...enormous public expending for treatment and rehabilitation of the affected individuals. The role of microRNAs in the pathophysiology of cerebral ischemia has been highlighted in current investigations. In addition, recent studies have also highlighted physical exercise as a possible protective factor both in the prevention and in the effects of cerebral ischemia, placing it as an important study resource. Thus, we investigated the role of physical exercise in experimental cerebral ischemia associated with the expression of microRNA-27b. 16 animals were used, divided into four experimental groups: Control, Physical Exercise, Cerebral Ischemia and Cerebral Ischemia associated with Physical Exercise. The real-time PCR methodology was used to analyze the expression of microRNA-27b. Although there were no statistically significant differences in the expression of microRNA-27b between the groups studied, the increased expression of microRNA-27b in the Physical Exercise group indicates its neuroprotective role in the pathophysiology of cerebral ischemia. KEY WORDS: Cerebral ischemia; Physical exercise; MicroRNAs; MicroRNA-27b.
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