The Association of Methylation Status and Expression Level of MyoD1 with DNMT1 Expression Level in Breast Cancer Patients.

Background: Breast cancer (BC) is the most common malignancy in women worldwide. The methylation status of MyoD1, a tumor suppressor gene, is enrolled in various cancers, i.e., BC. Various studies showed the impact of MyoD1 epigenetic dysregulation in BC. This study aimed to investigate the methylation status and expression level of MyoD1 in BC patients and its association with the expression of DNMT1. Materials and Methods: This case-control study was conducted on 30 cases (pathology-confirmed ductal carcinoma) and 18 controls (fibroadenoma and fibrocystic masses), referred to Velayat Hospital, Qazvin, Iran. The expression of the MyoD1 and DNMT1 and the promoter methylation of the MyoD1 were evaluated in tissue blocks of BC patient masses using qRT-PCR and MS-PCR assays, respectively. SPSS 24.0 was used to analyze the data. Results: The MyoD1 promoter is hypermethylated in BC patients compared to controls (p =0.001). The expression level of MyoD1 in BC patients was significantly reduced compared to controls (fold change =0.13, p =0.042). In addition, in BC patients, the reduced expression level of MyoD1 was significantly associated with methylation of the MyoD1 promoter (p =0.001). There is no significant difference between the expression level of DNMT1 in BC patients and controls (p =0.197). A significant association is found between the expression of DNMT1 and the methylation status of the MyoD1 promoter (p =0.038). Discussion: The expression level of MyoD1 is affected by the methylation status of the promoter of this gene. Moreover, the expression level and methylation status of MyoD1 are correlated with clinical parameters.

Evaluation of IGF2, KRT14, and KRT20 as Urinary Biomarkers in Patients with Bladder Cancer.

Background: Many researchers have tried to identify bladder cancer biomarkers to reduce the need for cystoscopy. The aim of this study was to identify and measure appropriate transcripts in patient urine to develop a non-invasive screening test. Methods: From February 2020 to May 2022, 49 samples were obtained from Velayat Hospital, Qazvin University of Medical Sciences, Qazvin, Iran. Twenty-two samples were obtained from bladder cancer patients and 27 from bladder cancer-free subjects. RNA was extracted from participant samples, quantitative RT-PCR was performed, and TNP plots were used to assess IGF2 (NCBI Gene ID: 3481), KRT14 (NCBI Gene ID: 3861) and KRT20 (NCBI Gene ID: 54474) expression. For UCSC Xena analysis, Dataset ID: TCGA-BLCA was used to compare transitional cell carcinoma (TCC) and normal samples for survival rates. Results: IGF and KRT14 were more greatly expressed in patient urine samples than in those of the normal group. However, KRT20 expression did not significantly differ between the two groups. IGF2 had 45.45 and 88.89% sensitivity and specificity, respectively, for detecting TCC in urine samples while KRT14 had 59 and 88.89% sensitivity and specificity, respectively. Also, these results infer that overexpression of IGF would be prognosticators of poor TCC outcomes. Conclusion: Our study showed that IGF2 and KRT14 are overexpressed in bladder cancer patient urine, and IGF2 could be a potential biomarker for poor prognoses in TCC.

Detection of bacterial agents causing prostate infection by culture and molecular methods from biopsy specimens.

Background and Objectives: Prostatitis affects about 16% of men in their lifetime and sometimes leading to prostate cancer. Bacterial infections are the most common causes of prostatitis. Diagnosis of the causative agents of bacterial prostate infections plays an essential role in timely treating and preventing secondary complications. This study isolated bacterial infectious agents in patients' surgical prostate and evaluated them by routine and molecular microbiological methods. Materials and Methods: In this cross-sectional study, 72 prostate biopsy specimens were collected from the Orology Departmen of hospitals of Qazvin University of Medical Sciences. All samples were cultured in aerobic and anaerobic conditions. Antibiotic susceptibility test by Kirby-Bauer standard method was performed for all isolated bacteria. In addition, all isolated bacteria were identified using 16S rDNA PCR and sanger sequencing methods. Also, TaqMan real-time PCR was applied to detect Ureaplasm aurealyticum, Mycoplasma hominins, and Mycoplasma genitalium. Results: In conventional culture method, out of 18 positive samples, 15 samples (83.3%) were Gram-negative bacteria and 3 samples (16.6%) were Gram-positive bacteria, containing Escherichia coli (55.5%), Klebsiella pneumoniae (11.1%), Enterobacter cloacae (5.5%), Pseudomonas aeruginosa (11.1%), Staphylococcus aureus (11.1%), and Enterococcus faecalis (5.5%). The results of molecular identification methods were the same as conventional culture results. Also, four patients were Ureaplasm aurealyticum, and three patients were positive for Mycoplasma hominis. Conclusion: Most bacteria isolated from prostate specimens belonged to the Enterobacteriaceae family, especially Escherichia coli, Klebsiella pneumoniae, Enterobacter cloacae. Staphylococcus aureus and Enterococcus faecalis were cocci isolated in the specimens too. Also, Ureaplasma urealyticum, and Mycoplasma hominis were identified in prostatitis.

Development of a SYBR Green Multiplex Real Time PCR for Simultaneous Detection of Mycobacterium Tuberculosis and Nocardia Asteroides in Respiratory Samples.

BACKGROUND: Nocardia asteroides and Mycobacterium tuberculosis are worldwide-distributed bacteria. These infectious agents can cause many infections in humans, especially in immunocompromised individuals. Pulmonary infections are more common and have similar clinical symptoms. Proper diagnosis and treatment of these patients are important for accurate treatment and could be lifesaving. METHODS: In this study, a multiplex real-time PCR assay was established for the simultaneous detection of the N. asteroides and M. tuberculosis. Both this homemade multiplex real time PCR and routine commercial tuberculosis tests were performed on 150 pulmonary specimens collected from individuals suspected to have tuberculosis. RESULTS: From 150 specimens, 20 samples were acid fast positive, 14 positives for M. tuberculosis by singleplex real time PCR, 10 positives for N. asteroides by singleplex real time PCR and 2 positives for M. tuberculosis and N. asteroides by multiplex real time PCR whereas 14 samples were positive for M. tuberculosis with commercial test. Differential diagnosis of pulmonary tuberculosis is useful for their proper treatment. CONCLUSION: Our test had good performance for differential diagnosis of tuberculosis and nocardiosis. Therefore, it is recommended to be used to diagnose such patients.

The relationship between gastric cancer and Helicobacter pylori in formaldehyde fixed paraffin embedded gastric tissues of gastric cancer patients-scorpion real-time PCR assay findings.

Gastric cancer is the second leading cause of cancer-related deaths worldwide and it seems that environmental and lifestyle factors and infection with Helicobacter pylori (H. pylori) have had a major role in the etiology of gastric cancer. The aim of this study was to investigate the presence of H. pylori DNA in archival gastric tissues of patients with gastric cancer disease by rapid, sensitive and specific technique of Scorpion Realtime PCR. This retrospective cross-sectional study was performed on 285 paraffin embedded gastric specimens of patients who were pathologically proved for gastric cancer admitted in Bou-Ali, Shahid Rajaie and Dehkhoda hospitals and Bahar and Farzam private laboratory in Qazvin city in Iran during 2009 and 150 paraffin embedded pathological specimens of patients with other proved diagnosis other than gastric cancer. Results of our Scorpion Realtime PCR analysis showed that DNA of H. pylori DNA was present in 78.42% of our total specimens. Modified McMullen's Staining of paraffin embedded sections was positive in 210 patients. Also we were not able to finding significant relationship between demographic characteristics of our studied patients and presence of H. pylori DNA in their formaldehyde fixed paraffin embedded gastric tissues samples. Existence of H. pylori in gastric tissue samples of patients with gastric cancer is controversial and our results indicated that in our studied specimens prevalence of H. pylori was significantly more than recent published reports.

Patho-epidemiological features of esophageal and gastric cancers in an endemic region: a 20-year retrospective study.

BACKGROUND: Gastric and esophageal cancers are among the most lethal human malignancies worldwide. Of all malignancies estimated in Iran (47,100), gastric and esophageal cancers were responsible for 7,800 and 3,500 deaths in 2008 respectively. The present study aimed to provide an image of patho-epidemiological characteristics with their trends during two past decades with emphasis on topographic, morphologic, and some demographic features. MATERIALS AND METHODS: In a hospital-based retrospective study in 2009, all pathological reports from esophageal endoscopies and gastric biopsies through a 20 years period (1989-2008) were collected and analyzed in four interval periods (five years each). Also, all eligible samples in hospital archives were enrolled for further testing. Besides, demography, topography and morphology of all samples were determined and analyzed by statistical software. RESULTS: No significant statistical difference was seen in frequency of esophageal and gastric tumors throughout the study. Esophageal cancer cases were older than gastric. Sex ratio was 2.33/1 and men had a higher rate of both esophageal and gastric tumors. Stomach cancer included 64.3% of all cases. Inferior third and end of esophagus were common locations for esophageal tumors whereas proximal stomach was common for gastric tumors. Squamous cell carcinoma and adenocarcinoma were common morphological types of tumors in esophagus and stomach respectively. CONCLUSIONS: Morphological trends showed an increase of esophageal adenocarcinoma and diffuse/intestinal ratio in stomach cancers. Trends in incidence from gastric cancer decreased based on topographic studies but we could not find a topographical trend toward cardia.

Relationship between bone mineral density and maturity index in cervical smears, serum estradiol levels and body mass index.

OBJECTIVES: Osteoporosis is a systemic skeletal disease with a consequent increased risk of fracture, decreased quality of life and economic burdens for both the patients and health care system. While Dual energy X-Ray absorptiometry remains the gold standard for assessment of bone mineral density, it cannot be requested for all patients for obvious reasons. By determining other variables that may correlate with osteoporosis, we can identify individuals who may be at risk for osteoporosis earlier. Then, they can be treated at the earlier stages of the disease. In the present study, relationships between bone mineral density, maturity index in cervical smear, serum estradiol level and body mass index were examined. MATERIALS & METHODS: The present study performed on 128 women, who had been referred for bone mineral densitometry. Blood samples were obtained for determination of serum estradiol level. Cervical smear was taken for assessment of cell's maturity. Cervical smears were examined by a pathologist and were sorted as atrophic or mature. Body mass index was calculated too. Relationships among Body mass index, serum estradiol level, and maturity index and bone mineral densitometry were analyzed using proper statistical tests. RESULTS: Maturity index had significant relationship with T Score in the spine and femoral neck (P<0.001). Significant relationships were found between serum estradiol level and femoral neck T Score (P<0.004) and spine T Score (P<0.008). Also a significant relationship was found between body mass index and bone mineral density. CONCLUSION: Pap smear is a routine examination which is performed mainly for screening purposes in gynecology. It is non-invasive, simple and low-priced. Results of current study suggest that women with atrophic cervical smear should be examined more rigorously for osteoporosis. If any patient has atrophic maturity index in her cervical smear, she will be at much higher risk of osteoporosis.