Proof of concept that melanoma nuclear count compares favourably with the benchmark histological prognostic feature, Breslow thickness.

AIMS: Breslow thickness (BT) is the most important histological prognostic feature for melanoma prognosis, but it only captures tumour size in one dimension. Adding a further measurement in a different axis has been shown to improve prognostic value. It seems reasonable that further prognostic value could be obtained by estimating the number of invasive melanoma cells using nuclear count. The aim of this study was to show proof of concept that nuclear count has prognostic value independent of BT. METHODS AND RESULTS: Melanoma cell nuclei were labelled with SRY-related HMG-box 10 (SOX10) protein, the sections scanned and StarDist machine-learning algorithm used to count nuclei in 102 cases of primary cutaneous melanoma. Prognostic value was assessed using survival analyses. Nuclear count correlated strongly with T category, BT and calculated tumour area (each P < 0.001), suggesting that it was a valid marker of melanoma burden. Nuclear count was a predictor for overall survival in univariable analysis [hazard ratio (HR) = 2.25, confidence interval (CI) = 1.66-3.06, P < 0.001] and multivariable analysis (HR = 2.60, CI = 1.59-4.24, P < 0.001). BT and ulceration were significant in univariable analyses, but not in multivariable models with nuclear count. Models containing nuclear count showed the best fit. Similar results were seen for melanoma-specific and metastasis-free survival. Nuclear count was able to stratify melanomas within a given T stage. CONCLUSIONS: This study demonstrated proof of concept that counting melanoma nuclei may be an improved measure of invasive tumour burden compared to BT. Future studies will need to refine methods of nuclear detection and also to confirm its prognostic value.

The High Mutational Sensitivity of ccdA Antitoxin Is Linked to Codon Optimality.

Deep mutational scanning studies suggest that synonymous mutations are typically silent and that most exposed, nonactive-site residues are tolerant to mutations. Here, we show that the ccdA antitoxin component of the Escherichia coli ccdAB toxin-antitoxin system is unusually sensitive to mutations when studied in the operonic context. A large fraction (∼80%) of single-codon mutations, including many synonymous mutations in the ccdA gene shows inactive phenotype, but they retain native-like binding affinity towards cognate toxin, CcdB. Therefore, the observed phenotypic effects are largely not due to alterations in protein structure/stability, consistent with a large region of CcdA being intrinsically disordered. E. coli codon preference and strength of ribosome-binding associated with translation of downstream ccdB gene are found to be major contributors of the observed ccdA mutant phenotypes. In select cases, proteomics studies reveal altered ratios of CcdA:CcdB protein levels in vivo, suggesting that the ccdA mutations likely alter relative translation efficiencies of the two genes in the operon. We extend these results by studying single-site synonymous mutations that lead to loss of function phenotypes in the relBE operon upon introduction of rarer codons. Thus, in their operonic context, genes are likely to be more sensitive to both synonymous and nonsynonymous point mutations than inferred previously.

Effect of Rose Syrup and Marigold Powder on the Physicochemical, Phytochemical, Sensorial and Storage Properties of Nutricereals and Milk-Based Functional Beverage.

ABSTACTAim: The present study aimed to investigate the effect of rose sirup and marigold powder on the physicochemical properties, bioactive potential, sensory acceptability and storage life of the nutricereals (finger millet, oats) and milk-based functional beverage (FB).Method: Preliminary trials were performed using different levels of rose sirup (8-14%) and marigold powder (0.40-0.55%) in the pre-standardized FB. The most acceptable concentration was selected on the basis of sensory analysis. Selected beverages were then subjected to the physicochemical analysis, assessment of bioactive compounds and FTIR characterization. The effect of flower extracts on the mineral content and storage life (4 ± 1 °C) of beverages was also studied. The significant difference in treatments was determined using Duncan's multiple range test, SPSS 25.0.Results: The best acceptable concentrations for rose sirup and marigold powder were 10% and 0.50%, respectively. A significant (p ≤ 0.05) decrease in the dietary fiber (6.50%) and ß-glucan (3.95%) content was observed on the addition of rose sirup. Significant (p ≤ 0.05) increase in the total phenols (119.18-145.23%), ß-carotene (0.37%), anthocyanins (78.82-230.58%) and antioxidant activity (4.98-7.17%) was observed on the addition of flower extracts. Strong peaks were observed in the regions of 3600-3200, 3000-2800 and 1700-1600 cm - 1 on FTIR characterization. A significant decrease in the mineral content of FB was also found on the addition of rose sirup. Rose flavored beverage had the highest overall acceptability (7.83 ± 0.23) and storage stability (50 days at refrigerated storage) among the prepared beverages.Conclusion: The addition of flower extracts significantly improved the acceptability of the prepared beverages. It not only improved the phytochemical profile but also had a substantial impact on storage stability.

Contribution of the Chromosomal ccdAB Operon to Bacterial Drug Tolerance.

One of the first identified and best-studied toxin-antitoxin (TA) systems in Escherichia coli is the F-plasmid-based CcdAB system. This system is involved in plasmid maintenance through postsegregational killing. More recently, ccdAB homologs have been found on the chromosome, including in pathogenic strains of E. coli and other bacteria. However, the functional role of chromosomal ccdAB genes, if any, has remained unclear. We show that both the native ccd operon of the E. coli O157 strain (ccdO157) and the ccd operon from the F plasmid (ccdF), when inserted on the E. coli chromosome, lead to protection from cell death under multiple antibiotic stress conditions through formation of persisters, with the O157 operon showing higher protection. While the plasmid-encoded CcdB toxin is a potent gyrase inhibitor and leads to bacterial cell death even under fully repressed conditions, the chromosomally encoded toxin leads to growth inhibition, except at high expression levels, where some cell death is seen. This was further confirmed by transiently activating the chromosomal ccd operon through overexpression of an active-site inactive mutant of F-plasmid-encoded CcdB. Both the ccdF and ccdO157 operons may share common mechanisms for activation under stress conditions, eventually leading to multidrug-tolerant persister cells. This study clearly demonstrates an important role for chromosomal ccd systems in bacterial persistence.IMPORTANCE A large number of free-living and pathogenic bacteria are known to harbor multiple toxin-antitoxin systems, on plasmids as well as on chromosomes. The F-plasmid CcdAB system has been extensively studied and is known to be involved in plasmid maintenance. However, little is known about the function of its chromosomal counterpart, found in several pathogenic E. coli strains. We show that the native chromosomal ccd operon of the E. coli O157 strain is involved in drug tolerance and confers protection from cell death under multiple antibiotic stress conditions. This has implications for generation of potential therapeutics that target these TA systems and has clinical significance because the presence of persisters in an antibiotic-treated population can lead to resuscitation of chronic infection and may contribute to failure of antibiotic treatment.

Molecular Determinants of Mutant Phenotypes, Inferred from Saturation Mutagenesis Data.

Understanding how mutations affect protein activity and organismal fitness is a major challenge. We used saturation mutagenesis combined with deep sequencing to determine mutational sensitivity scores for 1,664 single-site mutants of the 101 residue Escherichia coli cytotoxin, CcdB at seven different expression levels. Active-site residues could be distinguished from buried ones, based on their differential tolerance to aliphatic and charged amino acid substitutions. At nonactive-site positions, the average mutational tolerance correlated better with depth from the protein surface than with accessibility. Remarkably, similar results were observed for two other small proteins, PDZ domain (PSD95pdz3) and IgG-binding domain of protein G (GB1). Mutational sensitivity data obtained with CcdB were used to derive a procedure for predicting functional effects of mutations. Results compared favorably with those of two widely used computational predictors. In vitro characterization of 80 single, nonactive-site mutants of CcdB showed that activity in vivo correlates moderately with thermal stability and solubility. The inability to refold reversibly, as well as a decreased folding rate in vitro, is associated with decreased activity in vivo. Upon probing the effect of modulating expression of various proteases and chaperones on mutant phenotypes, most deleterious mutants showed an increased in vivo activity and solubility only upon over-expression of either Trigger factor or SecB ATP-independent chaperones. Collectively, these data suggest that folding kinetics rather than protein stability is the primary determinant of activity in vivo This study enhances our understanding of how mutations affect phenotype, as well as the ability to predict fitness effects of point mutations.

Challenges and Opportunities for Equity in US School Meal Programs: A Scoping Review of Qualitative Literature Regarding the COVID-19 Emergency.

The emergency school meals program provided free meals to children in the United States (US) during COVID-19-related school closures. This scoping review aims to synthesize the existing qualitative empirical evidence published between March 2020 and January 2023 on the operations and utilization of emergency school meal programs during the pandemic. Qualitative, US-based peer-reviewed literature was collected from three sources: (1) parent review of all federal nutrition assistance programs; (2) systematic search of four databases; and (3) manual search of grey literature. Identified scientific articles and grey literature reports (n = 183) were uploaded into Covidence and screened for duplicates and inclusion/exclusion criteria. Our final sample included 21 articles/reports, including 14 mixed methods and seven qualitative-only projects. Articles spanned all U.S. states. Articles had similar research questions to understand school meals and/or general food access during COVID-19, with an emphasis on long-term policy implications. Hybrid deductive/inductive analytic coding was used to analyze data, utilizing domains from the Getting to Equity Framework (GTE). GTE considers multi-level factors that influence nutrition behavior while centering more equitable pathways to improve nutrition security and reduce adverse health. Findings were sorted into two categories: operational challenges during the pandemic and solutions to address inequities in school meal distribution during and after the pandemic, particularly during school closures such as summers or future emergencies. Key challenges related to supply chain issues, safety, and balancing families' needs with limited staff capacity. Programs addressed equity by (a) reducing deterrents through federally issued waivers and increased communications which allowed the serving of meals by programs to families who previously did not have access, (b) building community capacity through collaborations and partnerships which allowed for increased distribution, and (c) preparing and distributing healthy options unless barriers in supply chain superseded the effort. This review highlights the importance of emergency school meal programs and provides insights into addressing challenges and promoting equity in future out-of-school times. These insights could be applied to policy and practice change to optimize program budgets, increase reach equitably, and improve access to nutritious meals among populations at highest risk for nutrition insecurity.

Study of Association of Serum Prolactin Levels with Insulin Resistance in Type 2 Diabetes Mellitus Patients.

Objective: Prolactin (PRL) regulates glucose metabolism and insulin sensitivity. The study aimed to evaluate the role of PRL in glucose homeostasis and its association with insulin resistance in patients with diabetes mellitus (DM). Methods and Materials: This cross-sectional, observational study included 100 patients (25-60 years) with T2DM. Primary information including demographics, anthropometric measurements, and biochemical measures (complete blood count, glucose parameters, liver and kidney function test, lipid profile, thyroid function test, serum fasting insulin levels, serum PRL levels) was collected. Results: A total of 100 patients, 50 men and 50 women (25 premenopausal and 25 postmenopausal), were enrolled in this study. The correlation between serum cholesterol and PRL was found to be statistically non-significant (P = 0.129) in men and significant (P = 0.041) in women. There was an inverse relationship between fasting plasma glucose and serum PRL levels in both men (r = -0.88; P < 0.0001) and women patients (r = -0.768; P < 0.0001). Negative correlation between postprandial plasma glucose and PRL was found to be statistically significant (r = -0.398; P = 0.048) in postmenopausal women. The comparison in both men and women indicated an inverse correlation between serum PRL and glycated haemoglobin levels. There was a significant negative correlation between homeostasis model assessment-estimated insulin resistance (HOMA-IR) and PRL levels in both men (r = -0.362; P = 0.039) and women patients (r = -0.362; P = 0.003). Homeostasis model assessment of ß cell function (HOMA-ß), which directly correlates with residual pancreatic beta cell function, was positively correlated with prolactin levels, irrespective of gender and menopausal status of female subjects. Conclusion: Serum PRL levels correlate with improved glycaemic control.

Effect of household processing on nutritional and antinutritional composition, mineral-mineral ratios, and functional properties of Colocasia leaves.

Colocasia leaves are high in nutrients and other phytochemicals but their utilization remains limited due to a lack of awareness. Higher content of anti-nutritional factors like oxalic and tannic acid in Colocasia leaves limit nutrient availability. In the present study, the effect of four household procedures viz. soaking (8-12 h), microwave heating (2-6 min), cooking (30-60 min), and blanching (1-3 min), followed by sun drying, was studied on the nutritional, antinutritional and functional properties of Colocasia leaves. A significant increase in crude fibre (25.7%-29.65%), and protein (4.33-15.6%) content was found in all the treatments except for the microwave treatment. A significant decrease in fat (5.7-31.4%), ash (20.34-28.22%), oxalic acid (27.07-35.32%), and tannic acid (up to 96%) was also found in various treatments. Among the minerals, a significant increase was reported for calcium (up to 16.38%), and iron (up to 5.9%). The highest mineral retention was found in soaked samples. The soaked and cooked samples also had a higher Ca: Mg ratio. A significant change in functional properties was also found. FTIR peaks suggested no significant qualitative effect occurred on phytochemical or physicochemical characteristics. Cluster analysis showed that cooking was second to soaking in terms of overall quality which were most comparable to the control. Cooking efficiently reduced the antinutritional factors, however, a significant loss of nutrients and functional properties was also observed. Therefore, the soaking of Colocasia leaves for 8-10 h is recommended as the best practice before their food applications.

Rare complications of chicken pox in immunocompetent children: Acute respiratory distress syndrome.

Chickenpox is a highly contagious disease caused by varicella zoster virus. Fever and papulovesicular rash are hallmarks of this disease. These manifestations are self-liming and complete recovery is seen in most cases. We report two cases of chickenpox infection where the rare and atypical manifestations of acute respiratory distress syndrome with pleural effusion were seen.

COVID-19 presenting as acute adominal pain: A case series.

The coronavirus infection presents primarily as a respiratory illness, however, extra-pulmonary manifestations are known to occur, including gastrointestinal manifestations. Hereby, we report three cases of the COVID-19 infection who presented with acute-onset abdominal pain during illness. All three patients had respiratory symptoms suggestive of COVID-19 and abdominal symptoms consistent with acute pancreatitis, acute cholecystitis, and acute appendicitis. All three patients improved in terms of acute abdominal pain; however, the overall clinical course, the three illnesses were variable because of differences in underlying organ involment and pathophysiology.

Determination of functional similarity of biosimilar H9P2S from an investigational CHO clone with Adalimumab.

Biosimilars, which are replicas of innovator pharmaceuticals, constitute the most significant share of biopharmaceutical products. These products are associated with structural and manufacturing complexities and are hence considered as similar to innovator drugs. Adalimumab is a monoclonal antibody that has been approved by the US FDA for blocking TNF-α. Adalimumab, also known as Humira, is preferred over other anti-TNF-α mAbs because of its lower immunogenicity and enhanced clinical efficacy. As cost-effective mAb development is still a challenging area, we developed an in-house stable CHO-K1 cell line for the production of recombinant monoclonal mAb against TNF-α. This clone yielded H9P2S, as a biosimilar against TNF-α, for which several functional assays were conducted to prove its biosimilarity to Adalimumab. Two batches of H9P2S and their subsequent dilutions were compared with Adalimumab. H9P2S and Adalimumab showed highly similar TNF-α binding and neutralizing activities, confirming the suitability of our clone for yielding biosimilar drugs. Supplementary Information: The online version contains supplementary material available at 10.1007/s13205-022-03384-z.

Evaluation of lipoprotein (a) [Lp(a)] and lipid abnormalities in patients with newly detected hypertension and its association with severity of hypertension.

Introduction: Hypertension remains the major preventable cause of cardiovascular disease (CVD). Lipoprotein (a) is seen to be associated with established essential hypertension and contributes to atherogenesis or to thrombogenesis or both. Aim: Correlation between lipoprotein (a) [Lp(a)] and lipid abnormalities in patients with newly detected hypertension and its association with severity of hypertension. Methods: It was a cross-sectional observational study carried out at PGIMER, DR. RML Hospital, New Delhi, India. Estimation of serum Lp (a) and lipid parameters along with routine laboratory investigations were carried out in 100 newly diagnosed cases with hypertension and compared with age and sex matched 50 healthy normotensive controls. Result: Amongst 100 cases the mean systolic and diastolic blood pressure was 160.68 ± 19.75 mmHg and 84.44 ± 4.32 mmHg respectively. The mean serum Lp (a) in cases was 34.03 ± 7.55 mg/dl as compared to 24.13 ± 4.41 mg/dl in controls (p < 0.0001). 62% of cases as compared to 12% of controls had elevated serum Lp (a) levels. Apart from that, the levels of Lp (a) and lipid parameters increased significantly with higher stage of disease (p < 0.0001). Approximately 8% of cases had left ventricular hypertrophy as compared to 1% of control. Similarly, 18% of cases had Non-alcoholic fatty liver disease as compared to 4% of controls. 5% of cases had retinopathy as compared to nil in controls. 4% of cases had microalbuminuria as compared to nil in controls. Conclusion: It was observed that newly detected hypertension is associated with major derangements of Lp (a) and lipid parameters. We also concluded that end organ involvement is significantly higher in newly detected hypertensives as compared to normotensive subjects and it was attributed to be due to lipid abnormalities observed in the group.

A Stable CHO K1 Cell Line for Producing Recombinant Monoclonal Antibody Against TNF-α.

Monoclonal antibodies (mAbs) are one of the most significant molecules in protein therapeutics. They are employed in the field of immunology, oncology and organ transplant. They have been also been employed for alleviating several bacterial and viral infections. Moreover, they have revolutionized the area of targeted therapy and improved the quality of treatments, as compared to other cytotoxic drugs and therapies. mAbs bind to specific molecules on the antigen and exhibit specificity towards that molecule, i.e. epitope. Thus, mAbs have immense opportunity to be explored for personalized therapy. The introduction of targeted mAb-based therapeutics has promoted many important scientific achievements in rheumatology. This has warranted additional investigations for developing newer mAb producing clones, to supplement the limited industrial production of certain mAb therapeutics. In this investigation, an integrative approach comprising optimized expression, selection and expansion was adopted to develop a mammalian cell line expressing mAb against TNF-α.The resulting stable clone is anticipated to serve as an economic alternative to the industrial clones, especially for research purposes. The clone was constructed for development of biosimilar of the highly valued therapeutic antibody, Humira.

Barriers, Enablers, and Possible Solutions for Student Wellness: A Qualitative Analysis of Student, Administrators, and Staff Perspectives.

BACKGROUND: Child well-being, which includes physical, mental, and social health, has a critical effect on academic performance. Schools face many challenges in meeting needs for child well-being. Schools in low-resource areas, including the Mississippi Delta, face greater challenges when meeting student needs. METHODS: Eleven focus groups with middle school students and 12 interviews with teachers, staff, and administration in one Mississippi Delta middle school were completed in 2019. The Social Ecological Model and Theory of Organizational Readiness for Change were used as a framework for the study. RESULTS: Focus group data with middle school students and interviews with teachers, staff, and administrators show that academic stressors play a major role in student well-being; yet, teachers, staff, and administrators face immense pressure to achieve high test scores. However, students seem to be encouraged by a positive school environment, which could help minimize some stressors. CONCLUSIONS: It seems that small improvements to the school environment could have a significant effect on student well-being, especially when school infrastructure and limited resources do not support comprehensive changes.

Cognitive determinants of weight control by dietary patterns among postmenopausal women with osteoporosis: An application of theory of planned behavior.

Background: In this study, we aimed to assess the cognitive determinants of weight control behaviors by dietary patterns among postmenopausal women with osteoporosis. Methods: This cross-sectional study, based on the theory of planned behavior (TPB) was conducted from July to December 2017 among 240 postmenopausal women with osteoporosis in Tabriz, Iran. A validated and reliable TPB-based instrument, namely Weight-CuRB, and the food frequency questionnaire (FFQ) were used. Results: The results of exploratory factor analysis (EFA) indicated three dietary patterns (total variance explained=24.44%); healthy (n=71), mixed (n=78), and western (n=91). In addition, food items consumed by participants were classified into twenty-two food groups for dietary pattern analysis. In the healthy and western dietary patterns, attitude (ß: 0.140, P <0.001) and subjective norms (SNs) (ß: 0.498, P <0.01) were the only predictors of weight control behavior, respectively. In the women with healthy and western patterns, the TPB-based variables altogether explained 11% and 16% of variations in the behavior, respectively. Among all patients, the TPB-based variables explained 12.2% of variations in weight control behavior, within which SNs were the only significant predictor of the behavior (ß=-0.199, P <0.01). Conclusion: Our findings highlighted the remarkable role of dietary patterns in the associations between weight control and its cognitive determinants. Dietary patterns should be considered while designing weight control educational interventions among women with osteoporosis. In such interventions, promoting SNs and perceived behavioral control (PBC) should be considered as the core strategies to promote the behaviors among the patients who follow an unhealthy diet.

Luminescent Europium(III) "Turn-On" Sensor for G-Series Chemical Warfare Simulants: A Mechanistic Investigation.

A thermodynamically stable terpyridine-dicarboxylate based emissive Eu(III) probe, [Eu(OH-TPDC)] (1), was designed as a time-resolved "turn-on" luminescent probe for the selective sensing of diethyl chlorophosphate (DCP), a G-series nerve agent simulant through modulation of sensitization of 5D0 → 7FJ transitions from Eu(III) via quenching of nonradiative vibrational energy transfer.

Smart snacks in universities: possibilities for university vending.

Background: The study goal was to evaluate the nutritional impact of a healthy snack intervention on a southern university campus. Methods: This quasi-experimental study was conducted during the fall 2017 semester weekly for 14 weeks in a large southern U.S. university. For the intervention, half of vending snacks in four campus residential halls (housing from 216 to 361 students) were substituted with snacks complying with federal Smart Snacks in School nutrition standards for K-12 schools. For analysis, data from the Nutrition Facts labels of 14 vending machines or from manufacturer's websites was collected by trained graduate and undergraduate researchers. Results: On average, for each Smart Snack sold, there was a statistically significant reduction of 99.38 calories (CI=42.32, 156.43), 4 g saturated fat (CI = 2.23, 5.75), and 10.06 g of sugar(CI=2.92, 17.20). An average reduction of 41.88 mg in sodium and an increase of 0.81g in fiber was also found, but was not statistically significant. There was a significant difference (t(16)=3.02, P < 0.025, 95% CI = 10.77, 55.79) between the Quality Score of Smart Snacks (M=59.13,SD= ± 36.50) and that of non-compliant snacks (M=25.85, SD= ± 24.72). Conclusion: The nutritional impact with even a 50% Smart Snack replacement is promising. Many available comparable snacks mimic the mouthfeel, taste, and appearance of their original full-fat, full-sodium, and full-sugar counterparts. Including healthier snack choices in vending machines may be a viable option for universities to transform the campus eating environment.

Dental Caries in Relation to Socioeconomic Factors of 6 and 12-year-old Schoolchildren of Paonta Sahib, Himachal Pradesh, India: An Epidemiological Study.

AIMS AND OBJECTIVES: Determination of prevalence of dental caries in 6 and 12-year-old schoolchildren, dental caries prevalence between government and private schoolchildren, and the influence of socioeconomic factors on dental caries on children of Paonta Sahib, Himachal Pradesh, India. MATERIALS AND METHODS: In the government and private schools at Paonta Sahib of Sirmour district, Himachal Pradesh, India, a cross-sectional study was carried out. The study population comprised schoolchildren of 6 and 12 years. A total of 1,004 children of both genders were surveyed for dental caries status. The decayed, missing, filled teeth (DMFT) index was used for clinical examination of caries. Prior collection of sociodemographic information was done in addition to information on oral health behavior by personal interview. RESULTS: In the present study, dental caries prevalence was found to be higher in children of 6 years of age (63.6%) than that of 12 years (53.6%) (p = 0.001). The mean DMFT was 2.47 ± 2.75 for 6-year-old children and 1.18 ± 1.33 for 12-year-old children. No significant difference was observed in dental caries prevalence among children of government and private school (p = 0.167). Moreover, dental caries frequency was highest in children belonging to upper-middle (38.14%) class followed by lower-middle (32.32%), upper-lower (25.42%), and upper class (4.12%). The obtained data were analyzed using the chi-square test and one-way ANOVA test. CONCLUSION: Dental caries prevalence was high in children of 6 years of age in the study. The dental caries frequency was more in upper-middle-class children. These results suggest that emphasis should be given to scientific monitoring, effective implementation, and evaluation of school-based oral health promotion and preventive programs to increase the dental awareness and oral hygiene practices, in Paonta Sahib, Sirmour district. HOW TO CITE THIS ARTICLE: Jindal L, Dua P, Mangla R, et al. Dental Caries in Relation to Socioeconomic Factors of 6 and 12-year-old Schoolchildren of Paonta Sahib, Himachal Pradesh, India: An Epidemiological Study. Int J Clin Pediatr Dent 2020;13(4):395-398.

Disseminated tuberculosis presenting as massive lower gastrointestinal bleeding.

Intestinal tuberculosis has varied clinical presentations and often requires high index of suspicion for clinching the diagnosis. We report a case of an 18-year-old male who presented with abrupt onset of massive lower gastrointestinal bleeding and was diagnosed to have ileocecal tuberculosis. This case signifies the need to consider the possibility of intestinal tuberculosis as a cause of severe hematochezia among other etiologies to prevent mortality and morbidity.

Vector-related stratagems for enhanced monoclonal antibody production in mammalian cells.

Provision of effective monoclonal antibody (mAb) based therapeutics requires careful combination of expression, selection and screening, in a suitable cell culture platform. The cell culture platform, in turn, requires high performance expression vectors for consistent and high production of mAb-based therapeutics. Further, the components of the vector need to be optimized via genetic engineering approaches, to exclude non-productive clones, reduce low secretion levels, eliminate improper functioning that affect mAb heavy chain and light chain ratio, glycosylation and aggregation, which in turn may lead to low effector functions, and to enhance productivity and post-translational processing of mAbs. This manuscript focusses on the various genetic elements that are modified and incorporated into the expression cassette, to have amplified constructs and stably expressing clones, with homogenous specificity and yield of mAbs. Production of antibodies of human origin is now possible with recombinant approaches like the gene editing tools, which enable targeted insertion of the genes coding for mAbs, using customized nucleases that enhance gene insertion. However, the ratio of light chain and heavy chain of mAb cDNA under expression, factors influencing the extent of their expression and the signal peptides for mAbs are the factors that create bottlenecks for production of mAbs, at the cellular level. This review article is focused on understanding these limitations, to overcome the challenges associated with them, and thus spur the development of mAb-based therapeutics.