جالب میں اور کوٹ لکھپت جیل
ڈاکٹر اسرار شاہ
لاہور میں دوست جالب میلے کا انعقاد کر رہے ہیں اور میں کاغذ اور قلم پکڑے اپنے ماضی میں کھو گیا دوستوں نے اصرار کیا کہ اسرار شاہ لکھو۔
میری دعا ہے کہ کوئی نیا ضیاء الحق پیدا نہ ہو اور مجھے عمرِ رفتہ میں لے جائے میںکالج سے نکلوں تو ایسی یونیورسٹی میں داخل ہو جائوں جہاں ڈاکٹر مبشر حسین ،میاں محمود علی قصوری رائو رشید،رضا کاظم ایڈووکیٹ چوہدری اعتزاز حسین ،جسٹس سعید حسن ،آئی اے رحمن ،پروفیسر امین مغل،چوہدری اصغر خادم ،رشید قریشی ،شعیب ہاشمی ،حمید اختر ،محمد علی ایکٹر اور حبیب جالبؔجیسے پروفیسر اور اساتذہ نظر بند ہوں نئی نسل نا واقف ہے کہ یہ تمام لوگ اپنی ذات میں ایک ادارہ تھے اور ان میں کچھ آج بھی حیات ہیں ۔
کوٹ لکھپت جیل بھی کیا جیل تھی ،جیل کے سپرنٹنڈنٹ نے جیل کی دیوار کے ساتھ شام کو واک کر نے کی اجازت دی ڈاکٹر مبشر صاحب جیل میں ’’ماں ‘‘کا کردارادا کر رہے تھے وہ جیل سے راشن لیتے اس کو پکواتے تمام لوگ چٹانوں پر بیٹھتے اور سب میں برابر تقسیم کرتے ۔صبح دس بجے سے لے کر دوپہرکے کھانے تک عبدا ﷲملک صاحب کے کمرے میں سٹڈی سرکل ہو تا اور آئی اے رحمن صاحب لیکچر دیتے اور تمام سر نگوں ہوتے ۔
حاجی رشید انور جن کا تعلق مزدور کسان پارٹی سے تھا کیا خوبصورت انسان تھے عمر کے اعتبار سے وہ میرے والد کی طرح تھے جسم میں سی آئی اے چونا منڈی کے تششدد کی دردیں موجود تھیں وہ صبح میرے جسم کو دباتے اور بچوں کی طرح پکارتے ہوئے اٹھاتے کہ ’’اسرار شاہ ‘‘اٹھ جائو سورج نکل آ یا...
BackgroundFertility Preservation is the process of saving or protecting a person’s ability to have children in the future. It is often considered for individuals to undergo medical treatments that may impact their fertility, such as chemotherapy. Hence, the current study is aimed to assess clinical practitioners’ knowledge, practice, and attitude toward fertility preservation among oncologists. MethodsA cross-sectional survey was carried out to identify the current knowledge, attitudes, and practices regarding fertility preservation among oncologists—a total of seventy-nine oncologists in Karachi working in different public and private sectors. The study was conducted between January to August 2022. The data was gathered using a self-designed questionnaire that was distributed via email. ResultsOne hundred and eighty oncologists were provided with the survey form via email. The response rate was 47.22% (n=85). Out of which, six questionnaires were excluded due to incomplete information. The total questionnaire analyzed was n=79, which included 58 (73.41%) males and 21 (16.59%) females. ConclusionThe results revealed that oncologists had a compromised knowledge regarding fertility preservation for cancer patients. Despite weak knowledge, most oncologists believe that more elaborative measures should be taken to overcome this issue. DOI: https: //doi. Org/10.59564/amrj/01.01/004
Proteases are an important group of industrial enzymes and require to break long-chain polypeptides. Conventionally, proteases are classified into six different groups. Among them, serine proteases are most widely studied. Right now, these enzymes are used as the catalyst in many different industries, including leather, poultry, waste management, and, chemical. Despite a lot of merits, the majority of naturally occurring enzymes cannot be used at industrial scale due to their incompatibility with industrial conditions. Moreover, the high enzyme production cost is another bottleneck to large-scale application of enzyme in industry. Protein engineering is an effective way to address these problems. There are two protein engineering approaches available, directed evolution and rational design. Later is preferred due to accelerated process and small mutants’ library size. Furthermore, this is an efficient approach to add desirable characteristics through in-depth analysis of structure. There are many studies in which this approach has been successfully applied to improve catalytic efficiency and thermostability of enzymes. Thus, by applying this approach, we can produce cheaper and effective enzymes for large-scale industrial use. In this work, serine peptidase from locally isolated Pseudomonas aeruginosa strain was identified. Later on, to make enzyme thermostable site-directed mutagenesis of non-catalytic was carried out to increase temperature resistance and activity. The protease producing bacteria were isolated from tanneries waste and screened for their proteolytic activity using casein as the substrate. The bacterial strain, showing high caseinolytic activity was selected. The type of the protease, optimum pH and temperature was identified using the crude extract of bacterial culture. The PMSF assay revealed that the enzyme was serine protease. The bacterial strain was identified using 16S rRNA sequencing. It came to know that our bacterial strain belonged to Pseudomonas aeruginosa and named Pseudomonas aeruginosa BMB1. The 16S rRNA sequence was deposited to NCBI nucleotide database under accession number KY285994. Primers were designed using homologous sequences to amplify and sequence the serine protease (SP) gene from the bacterial strain. The gene sequence was submitted to NCBI nucleotide database under accession no MH045598. The computational characterization of the gene sequence deduced 50 kDa molecular weight serine protease with 7.01 PI along with 25 amino acid residues long N-terminal signal peptide sequence. Moreover, it was also deduced that there are three domains in the serine protease, including N-terminal trypsin domain, and two C-terminal PDZ domains (PDZ1 and PDZ2). The SP gene was cloned and expressed in three different expression vectors, including pET32(a) with 6X His tag, pET32 with TRX tag and pGEX 6p-1 with GST tag with and without N-terminal signal peptide sequence. As expected, we could not get the soluble expression of the SP with the signal peptide. The highest expression of the SP was observed in pET32(a) with 6X His tag. The Ni+2 affinity column was used to purify the SP after expression in BL21(DE3). The purified serine protease was characterized. The residual activity analysis of serine protease showed the small half-life; therefore, the rational protein engineering was used to elevate thermostability and half-life of SP. We used I-TASSER and FireProt to predict the 3D structure and stabilizing substitutions in the protein structure respectively. FireProt predicted eight stabilizing substitutions using consensus and energy-based approaches. Only two substitutions A29G and V336I were proved to be stable among eight. There was the substantial increase in the half-life of A29G and V336I as compared to their template. Moreover, both mutants exhibited 5 oC increase in Tm value compared to wild-type in thermal denaturation CD analysis. Furthermore, MD simulation also showed the same type of trends in thermal stability of both mutants. Interestingly, along with thermal stability, there was also an increase in catalytic efficiency of both mutants. The Km values of both mutants were smaller than wild-type, indicating the strong binding of the substrate with enzymes. The molecular docking analysis also showed a similar type of results. The increase in thermostability and catalytic efficiency of the mutants was possibly due to stabilization of the oligomeric state and strong intra-molecular interactions.