Evaluation of the CRISPR/Cas9 directed mutant TP53 gene repairing effect in human prostate cancer cell line PC-3

Prostate cancer is a common health problem among men worldwide and most of these prostate cancer cases are related to a dysfunctional mutant Tumor Protein p53 (TP53) gene. However, the CRISPR/Cas9 system can be used for repairing of a dysfunctional mutant TP53 gene in combination with donor single-stranded oligodeoxynucleotide (ssODN) via cells' own homology-directed repair (HDR) mechanism. In this study, we aimed to evaluate the CRISPR/Cas9 repairing efficiency on TP53 414delC (p.K139fs*31) null mutation, located in the TP53 gene, of human prostate cancer cell line PC-3 in combination with ssODNs. According to the next-generation sequencing results, TP53 414delC mutation was repaired with an efficiency of 19.95% and 26.0% at the TP53 414delC position with ssODN1 and ssODN2 accompanied by sgRNA2 guided CRISPR/Cas9, respectively. Besides, qPCR and immunofluorescence analysis showed that PC-3 cells, the TP53 414delC mutation of which were repaired, expressed wild type p53 again. Also, significantly increased number of apoptotic cells, driven by the repaired TP53 gene were detected compared to the control cells by flow cytometry analysis. As a result, sgRNA2 guided CRISPR/Cas9 system accompanied by ssODN was shown to effectively repair the TP53 414delC gene region and inhibit the cell proliferation of PC-3 cells. Therefore, the effects of the TP53 414delC mutation repairment in PC-3 cells will be investigated in the in vivo models for tumor clearance analysis in the near future

Role of VPAC1 and VPAC2 receptors in the etiology of pregnancy rhinitis: an experimental study in rats

Introduction: Pregnancy rhinitis is a common sex hormone-related otorhinolaryngological disorder. There are some epidemiological and physiological studies on pregnancy rhinitis, but histopathological and biomolecular changes have not been studied thoroughly. Objectives: The receptors VPAC1 and VPAC2 are known for their roles in allergic rhinitis. On the other hand, activation of subclinical allergy has been suggested in the pathophysiology of pregnancy rhinitis. Therefore, we aimed to compare the physiological and gestational pattern of VPAC1 and VPAC2 expression in rat nasal mucosa. Methods: Twenty adult Wister albino female rats were enrolled into the study. Two groups constituted as 10 control (group A) and 10 pregnant (group B) rats. They were fed ad libitum and sheltered at room temperature (22°±2 °C). The rats were sacrificed at the 20th day of gestation by intraperitoneal injection of 400 mg/kg Na-pentobarbitone. Then, 10 − 15 mL of blood was taken, and samples were reserved for the detection of serum estradiol and progesterone levels by ELISA test. The nasal septum was resected and divided in half for immunohistochemical analyses and real time polymerase chain reaction testing of VPAC1 and VPAC2. Results: VPAC1 and VPAC2 were found to be in all layers of septal specimens, but the immunostaining of surface epithelium was more distinct in specimens of both groups. We demonstrated higher overall staining intensity in the pregnant group. PCR revealed significant increase in expression of VPAC1 (p = 0.023) and VPAC2 (p = 0.021) in pregnant group when compared with control group. In addition, we demonstrated upregulatory effect of estradiol and progesterone on the vasoactive intestinal peptide receptor expression. Conclusions: Gestational up-regulation of nasal VPAC1 and VPAC2 was shown both by PCR and immunohistochemical analysis. These findings support the hypothesis that PR is caused by the activation of subclinical allergy that is present before pregnancy

Evaluation of serum MicroRNA expression profiles in patients with panic disorder

BACKGROUND: Studies on the role of microRNAs (miRNA) in anxiety disorders are limited. We aimed to determine the availability of miRNAs as biomarkers in serum and to demonstrate the changes of miRNAs expression in patients with panic disorder (PD). METHODS: Thirty-five patients with PD and 35 healthy controls (HC) were evaluated with Structured Clinical Interview for DSM Disorders-I (SCID-I) and Panic Disorder Severity Scale (PDSS). In each group miRNA expression analysis was performed in venous blood by the Real-time Polymerase Chain Reaction (RT–PCR) method for genetic evaluation. RESULTS: Compared with the HC group, eight miRNA expression levels were found different in the PD group. Five of them were upregulated and three of them were downregulated. There was no correlation between the levels of miRNA expression with PDSS total score and PDSS sub-items. However, miR-1297 and miR-4465 expression levels were significantly different between the two groups. LIMITATIONS: There are some limitations in this research. Firstly the number of samples is small. Another limitation of our study is that the presence of medical illness and continuous drug use were not excluded when PD and HC groups were selected. CONCLUSIONS: Our research is the first miRNA expression study in patients with PD which excluded psychotropic use and additional psychiatric disorders. In the PD group, miR-1297 and miR-4465 expression was upregulated than compared to the HC group. miR-1297 and miR-4465 regulate the GABAA gene regions that affect GABAA receptor subtypes that thought to play a role in the aetiology of PD

USING 1,25(OH)2 VITAMIN D3 AND PARICALCITOL IN RATS, IN WHICH THE NEPHROTIC SYNDROME IS INDUCED BY PUROMYCIN (PAN), IT WAS EFFECT OF THE LEVEL OF PROTEINURIA, PODOCIN AND NEPHRIN MRNA WITH PCR

WOS: 00044399840036