CenC, a multidomain thermostable GH9 processive endoglucanase from Clostridium thermocellum: cloning, characterization and saccharification studies.

The growing demands of bioenergy has led to the emphasis on novel cellulases to improve efficiency of biodegradation process of plant biomass. Therefore, a thermostable cellulolytic gene (CenC) with 3675 bp was cloned from Clostridium thermocellum and over-expressed in Escherichia coli strain BL21 CodonPlus. It was attested that CenC belongs to glycoside hydrolase family 9 (GH9) with four binding domains, a processive endoglucanase. CenC was purified to homogeneity, producing a single band on SDS-PAGE corresponding to 137.11 kDa, by purification steps of heat treatment combined with ion-exchange chromatography. Purified enzyme displayed optimal activity at pH 6.0 and 70 °C. CenC had a half-life of 24 min at 74 °C, was stable up to 2 h at 60 °C and over a pH range of 5.5-7.5. Enzyme showed high affinity towards various substrates and processively released cellobiose from cellulosic substrates. It efficiently hydrolyzed carboxymethyl cellulose (30 U/mg), ß-Glucan Barley (94 U/mg); also showed activity towards p-nitrophenyl-ß-D-cellobioside (18 U/mg), birchwood xylan (19 U/mg), beechwood xylan (17.5 U/mg), avicel (9 U/mg), whatman filter paper (11 U/mg) and laminarin (3.3 U/mg). CenC exhibited Km, Vmax, Kcat, Vmax Km(-1) and Kcat Km(-1) of 7.14 mM, 52.4 µmol mg(-1) min(-1), 632.85 s(-1), 7.34 min(-1) and 88.63, respectively used CMC as substrate. Recombinant CenC saccharified pretreated wheat straw and bagasse to 5.12 and 7.31%, respectively at pH 7.0 and 45 °C after 2 h incubation. Its thermostability, high catalytic efficiency and independence of inhibitors make CenC enzyme an appropriate candidate for industrial applications and cost-effective saccharification process.

Breast Cancer Awareness and Associated Factors among Women in Pakistan: A Cross-Sectional Descriptive Study.

OBJECTIVE: Breast cancer is a highly prevalent cancer in females worldwide, with new cases around one million every year. In Pakistan, cancer of breast is the most common carcinoma in diagnosis, with one in nine females. Due to high burden of breast cancer in Pakistan, presented work purposes to investigate knowledge and awareness of breast carcinoma, its symptoms, and risk factors among Pakistani women, which play an important part in the early diagnosis of breast cancer. METHODS: A sample of 1000 females were approached generally from universities, hospitals, public places, local markets, rural areas and other cities for on-site data collection through face-to-face interviews and online data collection through telephonic interviews in Pakistan using the Breast Cancer Awareness Measure (BCAM). By using SPSS V. 25.0, the information  provided by the individuals first changed in awareness scores and was then analyzed. RESULTS: The study displayed, that mainstream participants lacked knowledge of breast carcinoma (63.2%) and the importance of its screening tools (64.7% and 83.2% lacked knowledge of mammography and BRCA tests respectively) for early detection. Almost 45% of respondents never noticed a change in their breasts. Most participants were unaware about breast cancer development is age-related and lifetime risk. Slightly more than 50% of the participants of the study were not knowledgeable regarding modifiable risk factors of breast carcinoma. A commonly known symptom was Breast lump by 53% of the respondents. Association was determined between demographic variables and breast cancer knowledge scores. Only 37.4% of respondents were found to be knowledgeable regarding breast cancer. CONCLUSION: BCAM is a productive instrument to assess awareness of breast carcinoma in females. The study indicated awareness of breast cancer is suboptimal in the population of Pakistan. Efforts should be made by public awareness campaigns and broadcasting of information about breast cancer and health education to increase awareness of risk factors.