Pyramiding and evaluation of segregating lines containing lectin and protease inhibitor genes for aphid resistance in Brassica juncea-Supplementary Data

Aphids are one of the most devastating pests, affecting the potential yield and quality ofBrassica juncea. In the current study, we have attempted to pyramid two transgenic lines containing chickpea lectin (CHPL, P1) and urdbean protease inhibitor (UPI, P2) in each under the phloem specific rolC promoter, through conventional breeding approach. In the derived F2 population, both lectin and protease inhibitor genes were segregating in a 9:3:3:1 ratio (p-value: 0.81), indicative of a single copy of the transgenes in the parents. Furthermore, the parental, as well as pyramided progenies were evaluated for their potential resistance to aphids in terms of mortality and natality. The lines containing both the transgenes were found to be superior over single gene transgenics as a higher mortality rate (96%) was found in F2on the 9th day as compared to single gene transgenics (86% and 80% in P1 and P2 respectively). A significant decrease in the number of nymphs was observed in P1 and P2 but most in F2 plants as almost 43, 32.08, and 107.5 times decrease in the number of nymphs was found in P1, P2, and F2 individuals over control. Expression profiling was done to see if there was any impact of gene pyramiding on the expression pattern of both transgenes before and after aphid treatment, and no significant changes were observed, indicating constitutive expression of transgenes in pyramided lines also. In conclusion, pyramided lines were found to be promising and were superior for aphid resistance.

Pyramiding and evaluation of segregating lines containing lectin and protease inhibitor genes for aphid resistance in Brassica juncea

Aphids are one of the most devastating pests, affecting the potential yield and quality ofBrassica juncea. In the current study, we have attempted to pyramid two transgenic lines containing chickpea lectin (CHPL, P1) and urdbean protease inhibitor (UPI, P2) in each under the phloem specific rolC promoter, through conventional breeding approach. In the derived F2 population, both lectin and protease inhibitor genes were segregating in a 9:3:3:1 ratio (p-value: 0.81), indicative of a single copy of the transgenes in the parents. Furthermore, the parental, as well as pyramided progenies were evaluated for their potential resistance to aphids in terms of mortality and natality. The lines containing both the transgenes were found to be superior over single gene transgenics as a higher mortality rate (96%) was found in F2on the 9th day as compared to single gene transgenics (86% and 80% in P1 and P2 respectively). A significant decrease in the number of nymphs was observed in P1 and P2 but most in F2 plants as almost 43, 32.08, and 107.5 times decrease in the number of nymphs was found in P1, P2, and F2 individuals over control. Expression profiling was done to see if there was any impact of gene pyramiding on the expression pattern of both transgenes before and after aphid treatment, and no significant changes were observed, indicating constitutive expression of transgenes in pyramided lines also. In conclusion, pyramided lines were found to be promising and were superior for aphid resistance.

Achievements of agricultural biotechnology: An initiative to double the farmer’s income through cutting edge technology

The population of the world is increasing nearly exponentially over time. To feed this population following the environment conservation protocol, it is essential to enhance the agricultural productivity even in the synchronizing agrarian land use pattern. To enhance the quality and productivity in agriculture sector, introducing the cutting edge technology is need of the hour.From ancient times, traditional approaches like selective breeding, adoption of agronomic management practices and application of indigenous technical knowledge have been used to attain resilience against various abiotic and biotic stresses. However, these traditional approaches are not sufficient to tackle the increasing repercussions of climate change and feed quality food to the expanding population. Therefore, in order to address these issues of climate change, population explosion and malnutrition, biotechnological interventions can be a promising approach. In the past, biotechnology based approaches have given successful products like Herbicide-resistant Soybean, Pusa Basmati 1, Bt Cotton, Bt Brinjal, Flavr-Savr tomato, a therapeutically significant product of Lithospermum erythrorhizon and Panax ginseng. Besides that many more need based products are in pipeline which is under scrutiny of regulatory bodies, policymakers and environmentalists. It is profoundly expected that in the coming day’s agricultural biotechnology applications will bring revolutionary changes to existing agricultural scenario. Therefore, in this review, we have summarized the achievement of agricultural biotechnology that is assisting to enhance the agricultural produce to double the income of farmers. However, this much is not enough; hence full utilization of all the sustainable agricultural biotechnological tools must come into the existence that definitely will boost the agricultural productivity.

Seroprevalence of Covid-19 infection among healthcare workers: A study from a tertiary care hospital in the National Capital Region of India

BACKGROUND Seroprevalence studies on severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection can provide information on the target populations for vaccination. We aimed to evaluate the seroprevalence among healthcare workers (HCWs) at our tertiary care institution and to identify parameters that may affect it. METHOD We assessed seroprevalence of SARS-CoV-2 by the chemiluminescence immunoassay test among 3258 HCW in our hospital and evaluated as per gender, age, their previous Covid-19 diagnosis, role in hospital and type/risk of exposure. RESULTS Of 3258 participants, 46.2% (95% CI 44.4%– 47.9%) were positive for SARS-CoV-2 IgG antibodies (i.e. IgG ?15 AU/ml). Higher seroprevalence was seen in non-clinical HCWs (50.2%) than in clinical HCWs (41.4%, p=0.0001). Furthermore, people with a history of Covid-19 were found to have significantly higher antibody levels (p=0.0001). Among the HCWs, doctors and nurses had lower relative risk (RR) of acquiring Covid-19 infection (RR=0.82; 95% CI 0.76–0.89) compared to non-clinical HCWs. CONCLUSION Seroprevalence in HCWs at our hospital was 46.2%. Clinical HCWs had lower seroprevalence compared to non-clinical HCWs. Previous history of Covid-19 almost doubled the seropositivity, particularly in those with current infection.

Safety, immunogenicity & effectiveness of the COVID-19 vaccine among healthcare workers in a tertiary care hospital

Background & objectives: The COVID-19 pandemic has caused significant global morbidity and mortality. As the vaccination was rolled out with prioritization on healthcare workers (HCWs), it was desirable to generate evidence on effectiveness of vaccine in prevailing real-life situation for policy planning. The objective of the study was to evaluate the safety, effectiveness and immunogenicity of COVID-19 vaccination among HCWs in a tertiary care hospital. Methods: This prospective observational study was undertaken on the safety, immunogenicity and effectiveness of the ChAdOx1 nCoV- 19 coronavirus vaccine (Recombinant) during the national vaccine roll out in January-March 2021, in a tertiary care hospital, New Delhi, India. Results: The vaccine was found to be safe, with local pain, fever and headache as the most common adverse events of milder nature which generally lasted for two days. The adverse events following vaccination were lower in the second dose as compared to the first dose. The vaccine was immunogenic, with seropositivity, which was 51 per cent before vaccination, increasing to 77 per cent after single dose and 98 per cent after two doses. Subgroup analysis indicated that those with the past history of COVID-19 attained seropositivity of 98 per cent even with single dose. The incidence of reverse transcription (RT)-PCR positive COVID-19 was significantly lower among vaccinated (11.7%) as compared to unvaccinated (22.2%). Seven cases of moderate COVID-19 needing hospitalization were seen in the unvaccinated and only one such in the vaccinated group. The difference was significant between the fully vaccinated (10.8%) and the partially vaccinated (12.7%). The hazard of COVID-19 infection was higher among male, age >50 yr and clinical role in the hospital. After adjustment for these factors, the hazard of COVID-19 infection among unvaccinated was 2.09 as compared to fully vaccinated. Vaccine effectiveness was 52.2 per cent in HCWs. Interpretation & conclusions: ChAdOx1 nCoV-19 coronavirus vaccine (Recombinant) was safe, immunogenic as well as showed effectiveness against the COVID-19 disease (CTRI/2021/01/030582).

Transcriptional Abundance of Myosin Light Chain 2 Gene in Cardiac Differentiated Canine Induced Pluripotent Stem Cells

Induced pluripotent stem cells (iPSCs) are promising cell source for cardiac tissue engineering and cell based therapies for heart repair as they can be expanded in vitro and differentiated into most cardiovascular cell types, including cardiomyocytes. During embryonic heart development, this differentiation occurs under the influence of internal and external stimuli that guide cells to go down the cardiac lineage. The aim of this study was to characterize the cardiac differentiation potential of a canine iPS cell. With the use of a standard embryoid body–based differentiation protocol for iPS cells were differentiated for 24 days. In vitro differentiations of canine iPSCs via embryoid bodies (EBs) were produced by ‘Hanging Drop’ method. EB’s were differentiated using 5-azacytidine (5-Aza). During differentiation, EBs were collected on day 4, 6, 8, 12, 16, 20 and 24 to evaluate the expression of cardiomyocyte specific marker. Analyses on molecular, structural, and functional levels demonstrated that iPS cell– derived cardiomyocytes show typical features of ES cell– derived cardiomyocytes. Reverse transcription polymerase chain reaction analyses demonstrated expression of marker genes. The differentiated cells expressed cardiac-specific gene myosin light chain 2 (MYL2) which started from day 8 of differentiation and highest expression was observed on day 16. Immunocytochemistry and relative expression of cardiac specific genes revealed that iPS cells differentiate into functional cardiomyocytes and allow to derivation of autologous functional cardiomyocytes for cellular cardiomyoplasty and myocardial tissue engineering

Perioperative Management of a Patient with Rheumatic Heart Disease Undergoing Emergency Laparotomy: A Case Report

Valvular heart disease and resulting comorbidity, such as heart failure or atrial fibrillation, significantly increase the risk for perioperative adverse events. Appropriate preoperative assessment, adequate perioperative monitoring, and early intervention may help to prevent adverse events and improve patient outcome.4 We are presenting a case report of 30 year old lady who was a known case of rheumatic heart disease and underwent emergency laparotomy for acute intestinal obstruction.

Study of Renal Functions in People Living with Human Immunodeficiency Virus/Acquired Immune Deficiency Syndrome (PLHA): An Observational Study of Tertiary Care Hospital in North India.

Aims: The aim of this study was to find out the involvement of renal dysfunction in people living with human immunodeficiency virus/acquired immune deficiency syndrome (PLHA) in India. Study Design: Observational study. Place and Duration of Study: Total (n=150) consecutive HIV positive patients between November 2012 - April 2014, who were attending the ART Clinic or were admitted in Department of Medicine at University College of Medical Sciences and Guru TegBahadur Hospital, Delhi, India were recruited for the study. Methodology: Estimated glomerular filtration rate (eGFR) was calculated by using Modification of Diet in Renal Disease (MDRD) and Chronic Kidney Disease Epidemiology Collaboration (CKD-EPI) equation. Morning spot urine samples were collected for urine albumin and urine creatinine test. Albumin/creatinine ratio (ACR) was calculated by using urine albumin and urine creatinine and were expressed in mg/g creatinine. Results: The mean eGFR (MDRD) of the study subjects was found 106.8±20.72 mL/min/1.73 m2 and a statistically significant difference was observed between male and female subjects (p = 0.039). The mean eGFR (CKD-EPI) of the study subjects was found 107.53±18.50 mL/min/1.73 m2, however, no significant difference was observed between male and female subjects (p = 0.745). The micro/macro-albuminuria (urinary ACR ≥ 30 mg/g creatinine) was found in 18 (12%) patients and leucocyturia and hematuria was found in 12.7% and 5% patients respectively. The median CD4 counts of the study subjects was 341 (222-467) cells/ mm3 and 141 (94%) were taking highly active anti-retroviral therapy (HAART). The most common HAART regimens were zidovudin/ lamivudine/nevirapine (ZLN) and tenofovir/ lamivudine/ nevirapine (TLN); 60.7% & 26.0% respectively. Hepatitis B and C co-infection rate found among subjects was 4% and 3.3% respectively. Conclusions: In conclusion, the prevalence of deranged renal functions as indicated by eGFR and urinary ACR is common in PLHA in North India. All HIV infected patients must undergo renal function tests including urinary ACR to detect the renal involvement at early stage.

Enzyme Based Asparagine Biosensor for the Detection of Asparagine Levels in Leukemic Samples.

Plants have been used for the treatment of different diseases since 1100 BC. Catharanthus roseus is one of the medicinally important plants showing anti-tumor, anti-mutagenic and anti-microbial effects. L-asparaginase is extracted by C. roseus and is used for the development of biosensors. L-asparaginase breaks down asparagine into aspartic acid and ammonia, thus killing the tumor cells due to asparagine starvation. In the present study the L-asparaginase was extracted from C. roseus and immobilized in different matrixes such as agar, soil, clay and k-carrageenan. Out of the various immobilization techniques k-carrageenan gave the fastest response time, so it was choosen for the development of asparagine biosensor. In the leukemic blood samples the asparagine concentration was 10-2 to 10-3 M whereas in normal blood samples it was 10-5 to 10-6 M. The developed biosensor gave the fast response and were sensitive and reliable.

Generation of transgenic mesenchymal stem cells expressing green fluorescent protein as reporter gene using no viral vector in caprine.

Mesenchymal stromal cells (MSC) are multipotent cells that can be derived from many different organs and tissues. While there are many ways to label and track cells each with strengths and weakness, the green fluorescent protein (GFP) is a reporter gene commonly employed. In the present study, caprine MSC were collected from bone marrow and cells were characterised with MSC specific markers. Passage 10 (P10) MSC cells were transfected using plasmid vector containing GFP as reporter gene with different concentrations of DNA and lipofectamine. Six different concentrations of DNA and lipofectamine as 1 µg DNA: 2 µL lipofectamine, 1 µg DNA: 2.5 µL lipofectamine, 1.2 µg DNA: 2.2 µL lipofectamine, 1.2 µg DNA: 2.5 µL lipofectamine, 1.5 µg DNA: 2.5 µL lipofectamine, 1.5 µg DNA: 3 µL lipofectamine were used. After 24 h and 48 h of transfection, caprine MSC were observed under florescent microscope. Highest transfection rate indicating green flourecscent MSC were found when the cells were transfected with 1.2 µg DNA: 2.2 µL lipofectamine and 1.5 µg DNA: 2.5 µL lipofectamine than other combinations. These cells have been propagated beyond 4th passage maintaining GFP expression. The results indicated that stable GFP positive MSC cells can be generated using the above protocol. These cells are being used for transplantation studies.

Common household spices: alternative source of antimicrobial compounds.

The antibacterial effect of some selected Indian medicinal plants was evaluated on bacterial strains like Bacillus cereus, Bacillus subtilis, Bacillus licheniformes and Escherichia coli. The solvents used for the extraction of plants were water, ethanol and methanol. The in vitro antibacterial activity was performed by agar well diffusion method. The most susceptible gram-positive bacteria was B. subtilis, The aqueous extracts of lemon was found to be effective against all the strains selected while aqueous extract of all other plant extract could not inhibit any of the bacterial strains investigated significantly. The most active antibacterial plant was Z. officinale Roscoe and Cinnamomum zeylanicum . The methanolic extracts were found to be more effective than etanolic and aqueous ones.