Antiproliferative Effect of 7-Ketositosterol in Breast and Liver Cancer Cells: Possible Impact on Ceramide, Extracellular Signal-Regulated Kinases, and Nuclear Factor Kappa B Signaling Pathways.

Background: This study aimed to examine the effect of 7-Ketositosterol (7-KSS), on sphingomyelin/ceramide metabolites and apoptosis in human breast MCF-7 and human liver HepG2 cancer cells. Methods: Anti-proliferative effects of 7-KSS treatment were assessed at different concentrations and periods. Cell viability was assessed through MTT analysis, whereas the levels of sphingosine-1-phosphate (S1P), sphingomyelins (SMs), and ceramides (CERs) were measured using LC-MS/MS. Phosphorylated 44/42 ERK1/2 and NF-κB p65 (Ser536) protein levels were measured by Western blot analysis and immunofluorescence staining. Apoptosis was evaluated by TUNEL staining and flow cytometric assessment of annexin-V and propidium iodide (PI) labeling. Results: Treatment with 7-KSS significantly decreased cell survival and S1P, p-44/42 ERK1/2, and p-NF-κB p65 protein levels in cancer cells compared to controls. A substantial rise was detected in intracellular amounts of C16-C24 CERs and apoptosis in cancer cells incubated with 7-KSS. Conclusions: 7-KSS stimulated ceramide accumulation and apoptosis while decreasing cell proliferation via downregulating S1P, p-44/42 ERK1/2, and p-NF-κB p65 protein levels.

Comparison of nylon-flocked swabs and cotton swabs in the detection of human papillomavirus infection in men.

BACKGROUND: Human papillomavirus (HPV) is an oncogenic virus and the commonest sexually transmitted pathogen worldwide. Appropriate sampling is an important factor in infection management. This study aimed to compare the efficacy of cotton swabs (CS) and nylon-flocked swabs (NFS) in sampling for HPV-DNA PCR testing in male patients with genital warts. METHODS: The study included men with genital warts who presented to the urology outpatient clinic of Antalya Medical Park Hospital. Before wart treatment, multisite sampling of the penis and genital area was performed separately with CS and NFS. The samples were analyzed for HPV-DNA using real-time PCR. RESULTS: The study included 45 men with a mean age of 32.1 ± 8.6 years. At least one HPV type was detected in all 45 patients with NFS sampling and 44 patients with CS sampling (total HPV types detected: 106 and 84, respectively). NFS sampling detected 52 high-risk HPV types in 37 of the 45 patients, while CS sampling detected 37 high-risk types in 19 patients (p = 0.029). NFS sampling also detected a total of 54 low-risk HPV types in all 45 patients, versus 47 low-risk HPV types in 41 patients with CS sampling. Multiple HPV types were detected in 30 patients with NFS and 17 patients with CS (p = 0.001). CONCLUSION: NFS were more effective than CS for HPV-DNA testing in men with genital warts. NFS were superior to CS in detecting multiple-type HPV infection and high-risk HPV types. The use of NFS should be recommended for HPV-DNA PCR testing in men.

Investigation of Mirna-223-3p Expression in Non-small Cell Lung Cancers.

BACKGROUND/AIM: Lung cancer remains a principal cause of cancer-related mortality worldwide. Non-small cell lung cancer (NSCLC), representing a significant 80-85% of lung cancer diagnoses, often presents at an advanced stage, with many patients demonstrating local growth or metastasis at the time of detection. Consequently, there exists a pressing need for augmented research into the molecular and genetic underpinnings of this malignancy to facilitate the development of innovative therapeutic and preventative strategies. MicroRNA's (miRNAs) are non-coding RNA molecules, consisting of 20-25 nucleotides. Their involvement in epigenetic processes holds a pivotal role in the elucidation of molecular mechanisms. This study examined the potential of miRNA-223-3p as a biomarker in the diagnosis of patients with NSCLC. MATERIALS AND METHODS: The expression analysis of miRNA-223-3p was performed in serum samples obtained from 18 patients diagnosed with NSCLC and a control group comprising 15 healthy volunteers. RESULTS: The miRNA-223-3p ΔCT values in the serum samples taken from the patient group exhibited statistically significant elevation compared to those of the control group (p=0.043). CONCLUSION: The present study corroborates previous literature indicating elevated levels of miRNA-223-3p in NSCLC cases and thereby suggesting its potential role as a biomarker in NSCLC cases.

Evaluation of the Relationship Between miRNA-22-3p and Gal-9 Levels in Glioblastoma.

BACKGROUND/AIM: Glioblastoma, the most prevalent primary malignant brain tumor, is significantly impacted by molecular mechanisms, including the function of microRNAs and galectins. The interplay between miRNA-22-3p and Galectin-9, a galactoside-binding lectin, is particularly notable. This study aimed to further investigate their roles in glioblastoma pathogenesis by analyzing the serum levels of these molecules in patients with glioblastoma. PATIENTS AND METHODS: This investigation included 50 subjects, consisting of 25 patients with glioblastoma and an equal number of healthy controls. Blood serum specimens were obtained for miRNA isolation and subsequent cDNA synthesis. The expression of the miRNA-22-3p gene was assessed using polymerase chain reaction (PCR), and a sandwich enzyme-linked immunosorbent assay (ELISA) was utilized to quantify serum Gal-9 concentrations. RESULTS: In patients diagnosed with glioblastoma, there was a significant elevation in miRNA-22-3p expression compared to healthy controls. However, despite a trend towards increased serum Gal-9 levels in the glioblastoma group, the difference did not reach statistical significance. CONCLUSION: Glioblastoma patients are characterized by increased Gal-9 serum levels and reduced miRNA-22-3p expression. These results indicate their potential as diagnostic and prognostic markers as well as therapeutic targets.

Effects of aurantiamide on a rat model of renovascular arterial hypertension.

Asperglaucide (ASP) is an aurantiamide, an effective constituent of purslane (Portulaca oleracea L.), a safe to eat greenery. Effects of ASP on endothelial function, endothelial nitric oxide synthase (eNOS) expression, vascular fluidity, renal and vascular reactive oxygen, and nitrogen species (ROS/RNS) production was examined in the two-kidney one-clip (2 K-1C) rat model of renovascular arterial hypertension. ASP toxicity, dose dependent eNOS gene expression and protein levels were also analyzed in human umbilical vein endothelial cells (HUVEC). The 2 K-1C model of hypertension was created via surgery and mean blood pressure (MBP) was measured by tail-cuff method during four weeks of ASP treatment. Erythrocyte deformability was monitored by rotational ektacytometry, while vascular constrictor and dilator responses were determined in organ baths. eNOS gene expression and protein levels were assessed in thoracic aorta and HUVEC. MBP was significantly decreased in hypertensive rats treated with ASP. Endothelium dependent vascular dilator and constrictor responses were also considerably improved following ASP treatment. There was a notable increase in red blood cell deformability in hypertensive rats treated with ASP as compared to hypertensive rats alone. A significant increase was observed in eNOS gene expression and protein levels in both normotensive and hypertensive rats treated with ASP. Treatment of HUVEC with 3 µM ASP notably increased eNOS mRNA and protein levels. In conclusion, ASP lowered blood pressure, improved endothelium-mediated relaxation, decreased renovascular ROS/RNS production in hypertensive rats. ASP also increased eNOS protein expression in aorta and HUVEC at nontoxic doses. ASP may have future potential as an anti-hypertensive agent.

Evaluation of the optimal sampling approach for HPV genotyping in circumcised heterosexual men with genital warts.

BACKGROUND: Human papillomavirus (HPV) causes a serious socioeconomic burden globally. However, there is currently no consensus on the optimal sampling method for HPVDNA genotyping in circumcised heterosexual men. This study aimed to determine the diagnostic efficacy of 6 different anatomic sampling sites in HPV DNA polymerase chain reaction (PCR) testing of circumcised heterosexual men with genital warts. METHODS: The study included circumcised heterosexual men who presented to our clinic with complaints of genital warts. Swab samples were obtained from the penile shaft (PS), scrotum, coronal sulcus (CS), and external urethral meatus (EUM). First-void urine (FVU) and genital wart biopsy (GWB) were also tested for HPV DNA by PCR. RESULTS: A total of 32 patients (mean age: 36.9 ± 6.9 years) were included. None of the six samples studied was sufficient on its own to reveal all HPV types detected in a patient. When the samples were analyzed individually, GWB detected an average of 49.5% of total HPV types in a patient. This rate was 50.5% for PS, 40.4% for CS, 31.6% for scrotum, 26.3% for EUM, and 15.8% for FVU samples. The detection rate increased to 75.8% with combined testing of GWB and PS samples, 83.2% with GWB/PS/CS, 90.5% with GWB/PS/CS/scrotum, and 98.9% with GWB/PS/CS/scrotum/EUM samples. CONCLUSION: No single anatomic region or sample type can detect all HPV types present in circumcised heterosexual men by PCR assay. The detection rate approaches 99% when wart biopsy is combined with swab sampling of the penile shaft, coronal sulcus, scrotum, and external urethral meatus.

Determination of IDH1, IDH2, MGMT, TERT and ATRX Gene Mutations in Glial Tumors.

BACKGROUND/AIM: The most frequent and dangerous kind of primary brain tumor is glioblastoma multiforme (GBM). The survival rates associated with GBM are very short and molecular markers for predicting survival are needed. The aim of our study was to evaluate isocitrate dehydrogenase 1 and 2 (IDH1, IDH2), telomerase reverse transcriptase (TERT), O-6- methylguanine-DNA methyltransferase (MGMT) and alpha-thalassemia/mental retardation, X-linked (ATRX) genes with next-generation sequencing (NGS) to find potential pathological mutations and their effect on survival. MATERIALS AND METHODS: Thirty patients who had undergone craniotomy and were diagnosed with high-grade glioma were evaluated for this study. Peripheral blood samples were obtained from all participants. IDH1, IDH2, TERT, MGMT and ATRX genes were evaluated with next-generation sequencing from the samples. Survival analysis evaluated the effects of all these mutations on survival. RESULTS: The median age of the patients was 58.5 (range=11- 74) years, and 56.7% (n=17) were under 60 years of age. According to sex, male patients comprised 66.7%. Targeted NGS detected 21 chromosomal aberrations. When more than three chromosomal anomalies were accepted as a reference, anomaly in three or fewer chromosomes negatively affected overall survival (hazard ratio=2.83). CONCLUSION: Targeted NGS generates therapeutically meaningful information, providing better prognostic information than conventional histology. Our study shows that NGS provides important information on survival by helping to detect chromosomal changes that can be detected in routine blood samples. It is clear that incorporating molecular diagnostics into our standard-of-care routine will help us better understand our patients' outcomes.

The Role of Kallikrein10 (KLK10) Polymorphism in Prostate Cancer Susceptibility.

PURPOSE: The present study aims to investigate the potential role of Kallikrein 10 (KLK10) genotype and allele frequencies in predisposition to prostate cancer. MATERIALS AND METHODS: KLK10 (rs7259451) gene polymorphisms were determined by real-time polymerase chain reaction analysis in patients with prostate cancer (n=69) and controls (n=76). RESULTS: KLK10 gene frequencies were significantly different in the case and control groups (P = .028). GG carriers were significantly higher in the control group (P = .034), whereas TT carriers were higher in the prostate cancer group (P = .033). Furthermore, The patients with GG genotype had the lowest PSA levels while TT carriers had the highest (P = .005). CONCLUSION:  According to the results, we suggested that carrying variant T allele and also carrying homozygote TT genotype could be a potential risk, while ancestral homozygote GG genotype and G allele are risk reducing factors for prostate cancer.

Investigation of Catechol-O-methyltransferase (COMT) gene Val158Met polymorphism in ovarian cancer

Objective: Catechol-O-methyltransferase (COMT), the product of the COMT gene, detoxifies the carcinogenic catechol estrogens. The aim of the present study was to examine the relationship between COMT Val158Met polymorphism and the risk of ovarian cancer. Material and Methods: The study groups consist of 94 individuals as a patients group with ovarian cancer (n=47) and control group (n=47). The allele and genotype frequencies were determined according to Hardy-Weinberg equilibrium (HWE). The allele and genotype frequencies. determined according to HWE. Genetic analysis were performed by real-time-polymerase chain reaction instrument, and the statistical analysis were performed by SPSS program. Results: Although no significant relationship was obtained among groups (p=0.413) regarding COMT gene Val158Met polymorphism, the genotype frequencies for COMT Val158Met (rs4860) polymorphism in groups was homozygote wild type GG genotype 25.5%, heterozygote GA genotype 46.8%, homozygote mutant AA genotype 27.7%. Conclusion: This study is the first to investigate the relationship between ovarian cancer and the Val158Met polymorphism in the COMT gene in a Turkish population. No statistically significant relationship was identified among genotypes belonging to the patient and control groups although sample sizes were relatively small and the analysis should be repeated in a larger cohort.