PURPOSE: The promoter methylation status of cell cycle regulatory genes plays a crucial role in the regulation of the eukaryotic cell cycle. CpG cytosines are actively subjected to methylation during tumorigenesis, resulting in gain/loss of function. E2F5 gene has growth repressive activities; various studies suggest its involvement in tumorigenesis. This study aims to investigate the epigenetic regulation of E2F5 in breast cancer to better understand tumor biology. METHODS: The promoter methylation status of 50 breast tumor tissues and adjacent normal control tissues was analyzed. mRNA expression was determined using SYBR® green quantitative polymerase chain reaction (PCR), and methylation-specific PCR was performed for bisulfite-modified genomic DNA using E2F5-specific primers to assess promoter methylation. Data was statistically analyzed. RESULTS: Significant (p<0.001) upregulation was observed in E2F5 expression among tumor tissues, relative to the control group. These samples were hypo-methylated at the E2F5 promoter region in the tumor tissues, compared to the control. Change in the methylation status (Δmeth) was significantly lower (p=0.022) in the tumor samples, indicating possible involvement in tumorigenesis. Patients at the postmenopausal stage showed higher methylation (75%) than those at the premenopausal stage (23.1%). Interestingly, methylation levels gradually increased from the early to the advanced stages of the disease (p<0.001), which suggests a putative role of E2F5 methylation in disease progression that can significantly modulate tumor biology at more advanced stage and at postmenopausal age (Pearson's r=0.99 and 0.86, respectively). Among tissues with different histological status, methylation frequency was higher in invasive lobular carcinoma (80.0%), followed by invasive ductal carcinoma (46.7%) and ductal carcinoma in situ (20.0%). CONCLUSION: Methylation is an important epigenetic factor that might be involved in the upregulation of E2F5 gene in tumor tissues, which can be used as a prognostic marker for breast cancer.
Biology ; Breast Neoplasms* ; Breast* ; Carcinogenesis ; Carcinoma, Ductal ; Carcinoma, Intraductal, Noninfiltrating ; Carcinoma, Lobular ; Cell Cycle ; Disease Progression ; DNA ; E2F5 Transcription Factor ; Epigenomics ; Eukaryotic Cells ; Genes, Regulator ; Humans ; Methylation* ; Polymerase Chain Reaction ; Promoter Regions, Genetic ; RNA, Messenger ; Up-Regulation*

Aims: Thermophilic proteases are important industrial enzymes because they can be used at high temperatures in various bioprocessing schemes. The bacterial population of the Cholistan desert was explored for thermophilic proteases and their industrial applications. Methodology and results: Three bacterial isolates K1, K5 and K7 were found promising protease producers. These isolates were preliminary identified as Bacillus based on morphological characteristics and biochemical tests (positive for catalase, oxidase and citrate tests, and negative for indole and urease tests). The isolates K1, K5 and K7 were further identified as Priestia endophytica, Lysinibacillus cresolivorans and Bacillus subtilis, respectively by phylogenetic analysis. The isolates grew best at 50 °C and P. endophytica (K1), L. cresolivorans (K5) and B. subtilis (K7) produced larger zones of hydrolysis at 37 °C, 45 °C and 50 °C at pH 7, respectively. The optimum temperature where protease activity was maximum was 65 °C for P. endophytica and L. cresolivorans and 55 °C for B. subtilis, and the optimum pH was 9. Conclusion, significance and impact of study The proteases produced by these isolates were found active at high temperatures (45 °C to 85 °C) and high pH (9-12), which make them industrially important thermoalkaliphilic proteases. These proteases successfully de-haired cow’s skin and de-stained blood from cotton cloth pieces, which are rarely tested applications of these proteases.
Desert ; Proteases ; Purification ; Thermophilic bacteria

Objective: To gather the fragmented literature on ethnobotany, phytochemistry and in-vitro activities of medicinal plants of Pakistan being used against common infections caused by Staphylococcus aureus (S. aureus). Methods: A large number of published and unpublished research studies related to the ethnomedicinal, phytochemical and anti-S. aureus activity of medicinal flora of Pakistan published from 1990-2018 were reviewed using online bibliographic databases such as PubMed, Web of Science, Science Direct, ResearchGate and libraries. Results: S. aureus can cause many human ailments including endocarditis, staphylococcal scalded skin syndrome, septic arthritis, respiratory problems with an estimated infection rate of 25%-35% across the globe. This review comprised of 86 medicinal plants. Data showed that people mostly used leaves (50%) for the preparation of traditional medicines. Correlation analysis on the reviewed data revealed that methanolic extract concentrations of medicinal plants was highly significantly positive correlated (r=0.8; P<0.01) with the S. aureus zone of inhibitions. S. aureus reportedly showed complete resistant to the commonly used antibiotic erythromycin. Isolated compounds like altheahexacosanyl lactone, cinnamaldehyde, niloticane, gobicusin A, asparacosin A, muzanzagenin, isoagatharesinol, friedelin, inophynone and eugenol were active against S. aureus. This study provided in-vitro proof for the flora of Pakistan used against different infections caused by S. aureus. Conclusions: Antibacterial agents from natural sources could be more effective against bacterial pathogens and will be helpful in minimizing the adverse effects of synthetic drugs, and hence provides a base for the pharmaceutical industries. http://www.apjtm.org/article.asp?issn=1995-7645;year=2018;volume=11;issue=6;spage=355;epage=368;aulast=Adnan;type=2.