Current achievements and future prospects of genetic engineering in Indian mustard (Brassica juncea L. Czern & Coss.).

MAIN CONCLUSION: Transgenic technology in Indian mustard has expedited crop improvement programs. Further, there is a need to optimize gene editing protocols and find out the suitable target genes to harvest the benefits of gene editing technology in this important edible oilseed crop. Brassica juncea is an economically and industrially important oilseed crop being grown mainly in India and in some parts of Canada, Russia, China and Australia. Besides being consumed as edible oil, it also has numerous applications in food and paint industry. However, its overall production and productivity are being hampered by a number of biotic and abiotic stress factors. Further, its oil and seedmeal quality needs to be improved for increasing food as well as feed value. However, the lack of resistant crossable germplasm or varieties necessitated the use of genetic engineering interventions in Indian mustard crop improvement. A number of genes conferring resistance to biotic stresses including lectins for aphids' control, chitinase, glucanase and osmotin for disease control and for abiotic stresses, CODA, LEA and ion antiporter genes have been transferred to Indian mustard. Both antisense and RNAi technologies have been employed for improving oil and seedmeal quality. Efforts have been made to improve the phytoremediation potential of this crop through genetic engineering approach. The deployment of barnase/barstar gene system for developing male sterile and restorer lines has really expedited hybrid development programs in Indian mustard. Further, there is a need to optimize gene editing protocols and to find out suitable target genes for gene editing in this crop. In this review paper, authors have attempted to review various genetic transformation efforts carried out in Indian mustard for its improvement to combat biotic and abiotic stress challenges, quality improvement and hybrid development.

Genetic diversity in leafy mustard (Brassica juncea var. rugosa) as revealed by agro-morphological traits and SSR markers.

Leafy mustard (B. juncea var. rugosa) constitutes an important group of vegetable mustard crops in India and is mainly cultivated in home-backyard and hilly regions of Uttarakhand and some North-eastern states. In the present study, various agro-morphological traits, physiological and biochemical traits along with SSR markers were used for genetic diversity evaluation in a germplasm collection of leafy mustard. This study revealed a significant variation among 59 accessions of leafy mustard in both qualitative and quantitative agro-morphological traits indicating the accessions' promising potential for consumption purpose and for use in breeding programs. Maximum variability was recorded for leaf area elongation rate (CV = 53.12%), followed by total plant weight (TPW) (CV = 50.63%) and seed yield per plant (CV = 44.33%). In molecular analysis, 155 SSRs evaluated resulted in 482 alleles and the number of alleles varied form 1 to 8 with an average of 3.11 alleles per marker. A total of 122 (78.70%) SSRs resulted into polymorphic amplicons. PIC value varied from 0.32 to 0.77 with an average value of 0.44 per SSR locus. The unweighted neighbour-joining-based dendrogram analysis divided all the 59 accessions into two major groups on the basis of both agro-morphological traits and SSR markers, whereas, three subpopulations/subgroups were predicted by population STRUCTURE analysis. AMOVA indicated the presence of more variability within population than among population. Overall, agro-morphologically better performing and genetically diverse genotypes have been identified which could be further used as donors for leafy mustard improvement programs.

Qualitative and quantitative reservoir characterization using seismic inversion based on particle swarm optimization and genetic algorithm: a comparative case study.

Accurate reservoir characterization is necessary to effectively monitor, manage, and increase production. A seismic inversion methodology using a genetic algorithm (GA) and particle swarm optimization (PSO) technique is proposed in this study to characterize the reservoir both qualitatively and quantitatively. It is usually difficult and expensive to map deeper reservoirs in exploratory operations when using conventional approaches for reservoir characterization hence inversion based on advanced technique (GA and PSO) is proposed in this study. The main goal is to use GA and PSO to significantly lower the fitness (error) function between real seismic data and modeled synthetic data, which will allow us to estimate subsurface properties and accurately characterize the reservoir. Both techniques estimate subsurface properties in a comparable manner. Consequently, a qualitative and quantitative comparison is conducted between these two algorithms. Using two synthetic data and one real data from the Blackfoot field in Canada, the study examined subsurface acoustic impedance and porosity in the inter-well zone. Porosity and acoustic impedance are layer features, but seismic data is an interface property, hence these characteristics provide more useful and applicable reservoir information. The inverted results aid in the understanding of seismic data by providing incredibly high-resolution images of the subsurface. Both the GA and the PSO algorithms deliver outstanding results for both simulated and real data. The inverted section accurately delineated a high porosity zone ( > 20 % ) that supported the high seismic amplitude anomaly by having a low acoustic impedance (6000-8500 m/s ∗ g/cc). This unusual zone is categorized as a reservoir (sand channel) and is located in the 1040-1065 ms time range. In this inversion process, after 400 iterations, the fitness error falls from 1 to 0.88 using GA optimization, compared to 1 to 0.25 using PSO. The convergence time for GA is 670,680 s, but the convergence time for PSO optimization is 356,400 s, showing that the former requires 88 % more time than the latter.

Envirotype-based delineation of environmental effects and genotype × environment interactions in Indian soybean (Glycine max, L.).

Soybean is a rainfed crop grown across a wide range of environments in India. Its grain yield is a complex trait governed by many minor genes and influenced by environmental effects and genotype × environment interactions. In the current investigation, grain yield data of different sets of 41, 30 and 48 soybean genotypes evaluated during 2019, 2020 and 2021, respectively across 19 locations and twenty years' data on 19 different climatic parameters at these locations was used to study the environmental effects on grain yield, to understand the genotype × environment interactions and to identify the mega-environments. Through analysis of variance (ANOVA), it was found that predominant portion of the variation was explained by environmental effects (E) (53.89, 54.86 and 60.56% during 2019, 2020 and 2021, respectively), followed by genotype × environment interactions (GEI) (31.29, 33.72 and 28.82% during 2019, 2020 and 2021, respectively). Principal Component Analysis (PCA) revealed that grain yield was positively associated with RH (Relative humidity at 2 m height), FRUE (Effect of temperature on radiation use efficiency), WSM (Wind speed at 2 m height) and RTA (Global solar radiation based on latitude and Julian day) and negatively associated with VPD (Deficit of vapour pressure), Trange (Daily temperature range), ETP (Evapotranspiration), SW (Insolation incident on a horizontal surface), n (Actual duration of sunshine) and N (Daylight hours). Identification of mega-environments is critical in enhancing the selection gain, productivity and varietal recommendation. Through envirotyping and genotype main effect plus genotype by environment interaction (GGE) biplot methods, nineteen locations across India were grouped into four mega-environments (MEs). ME1 included five locations viz., Bengaluru, Pune, Dharwad, Kasbe Digraj and Umiam. Eight locations-Anand, Amreli, Lokbharti, Bidar, Parbhani, Ranchi, Bhawanipatna and Raipur were included in ME2. Kota and Morena constitutes ME3, while Palampur, Imphal, Mojhera and Almora were included in ME4. Locations Imphal, Bidar and Raipur were found to be both discriminative and representative; these test locations can be utilized in developing wider adaptable soybean cultivars. Pune and Amreli were found to be high-yielding locations and can be used in large scale breeder seed production.

Utilization of crop wild relatives for biotic and abiotic stress management in Indian mustard [Brassica juncea (L.) Czern. & Coss.].

Brassica juncea (L.) Czern. & Coss. (Indian mustard) is an economically important edible oil crop. Over the years, plant breeders have developed many elite varieties of B. juncea with better yield traits, but research work on the introgression of stress resilience traits has largely been lagging due to scarcity of resistant donors. Crop wild relatives (CWRs) are the weedy relatives of domesticated plant species which are left unutilized in their natural habitat due to the presence of certain undesirable alleles which hamper their yield potential, and thus, their further domestication. CWRs of B. juncea namely include Sinapis alba L. (White mustard), B. tournefortii Gouan. (African mustard), B. fruticulosa Cirillo (Twiggy turnip), Camelina sativa L. (Gold-of-pleasure), Diplotaxis tenuisiliqua Delile (Wall rocket), D. erucoides L. (White wall rocket), D. muralis L. (Annual wall rocket), Crambe abyssinica R.E.Fr. (Abyssinian mustard), Erucastrum gallicum Willd. (Common dogmustard), E. cardaminoides Webb ex Christ (Dogmustard), Capsella bursa-pastoris L. (Shepherds purse), Lepidium sativum L. (Garden Cress) etc. These CWRs have withstood several regimes of biotic and abiotic stresses over the past thousands of years which led them to accumulate many useful alleles contributing in resistance against various environmental stresses. Thus, CWRs could serve as resourceful gene pools for introgression of stress resilience traits into Indian mustard. This review summarizes research work on the introgression of resistance against Sclerotinia stem rot (caused by Sclerotinia sclerotiorum), Alternaria blight (caused by Alternaria brassicae), white rust (caused by Albugo candida), aphid attack, drought and high temperature from CWRs into B. juncea. However, various pre- and post-fertilization barriers due to different ploidy levels are major stumbling blocks in the success of such programmes, therefore, we also insightfully discuss how the advances made in -omics technology could be helpful in assisting various breeding programmes aiming at improvisation of stress resilience traits in B. juncea.

Agro-morphological traits and SSR markers reveal genetic variations in germplasm accessions of Indian mustard - An industrially important oilseed crop.

Indian mustard is an economic and highly important industrial oilseed crop. In this study, genetic diversity among 135 Indian mustard germplasm accessions was evaluated using 11 agro-morphological descriptors and 227 SSRs. Morphological characterization of Indian mustard germplasm accessions exhibited a broad range of variation for characters including biological yield (CV = 25.63%), seed yield (CV = 23.23% and 1000-seed weight (CV = 23.14%); whereas traits such as days to maturity (CV = 2.91%) showed lowest degree of variation. Out of 227 SSR markers evaluated, a total of 159 (70.04%) SSRs produced polymorphic products and 68 (29.96%) SSRs resulted into monomorphic amplicons. The polymorphic markers amplified 575 alleles and the number of alleles ranged from 2-7 with 3.61 average number of alleles per locus. SSR markers BRMS-030, Ra2-E11, Ra2-G05, Ni4-G10 and Ol10B11 generated the highest number of alleles (7). SSR marker Ra2-G05 was having the highest allele frequency (0.84), while BRMS-002 was having the lowest major allele frequency (0.33). Polymorphism information content (PIC) values ranged from 0.24-0.61 with an average value of 0.39 per primer pair. Unweighted pair group method with arithmetic mean (UPGMA) cluster analysis based on morphological traits grouped Indian mustard genotypes into three clusters, while two clusters were obtained based on SSR based clustering. Population structure analysis provided a better estimate of genetic diversity and divided all the genotypes into five subpopulations. Genetically diverse accessions identified may be used for hybridization in Indian mustard crop improvement programs in future.

Molecular characterization and genetic diversity analysis in Indian mustard (Brassica juncea L. Czern & Coss.) varieties using SSR markers.

In this study, we evaluated genetic diversity in a panel of 87 Indian mustard varieties using 200 genomic-SSR markers. A total of 189 SSRs resulted into positive amplification with 174 (92.06%) SSRs generating polymorphic products and 15 (7.94%) SSRs producing monomorphic amplicons. A total of 552 alleles were obtained and allele number varied from 2-6 with an average number of 3.17 alleles per SSR marker. The major allele frequency ranged from 0.29 (ENA23) to 0.92 (BrgMS841) with an average value of 0.58 per SSR locus. The polymorphic information content (PIC) value ranged from 0.10 (BrgMS841) to 0.68 (BrgMS519) with 0.39 as mean PIC value. The gene diversity per locus ranged from 0.13 (BrgMS841) to 0.72 (ENA23 & BrgMS519) with a mean value of 0.48 per SSR primer pair. Both Unweighted Neighbor Joining-based dendrogram and population structure analysis divided all the 87 varieties into two major groups/subpopulations. Analysis of molecular variance (AMOVA) inferred the presence of more genetic variation (98%) among individuals than among groups (2%). A total of 31 SSRs produced 36 unique alleles for 27 varieties which will serve as unique DNA-fingerprints for the identification and legal protection of these varieties. Further, the results obtained provided a deeper insight into the genetic structure of Indian mustard varieties in India and will assist in formulating future breeding strategies aimed at Indian mustard genetic improvement.

Psychological distress and burnout among healthcare worker during COVID-19 pandemic in India-A cross-sectional study.

BACKGROUND: COVID-19 has inundated the entire world disrupting the lives of millions of people. The pandemic has stressed the healthcare system of India impacting the psychological status and functioning of health care workers. The aim of this study is to determine the burnout levels and factors associated with the risk of psychological distress among healthcare workers (HCW) engaged in the management of COVID 19 in India. METHODS: A cross-sectional study was conducted from 1 September 2020 to 30 November 2020 by telephonic interviews using a web-based Google form. Health facilities and community centres from 12 cities located in 10 states were selected for data collection. Data on socio-demographic and occupation-related variables like age, sex, type of family, income, type of occupation, hours of work and income were obtained was obtained from 967 participants, including doctors, nurses, ambulance drivers, emergency response teams, lab personnel, and others directly involved in COVID 19 patient care. Levels of psychological distress was assessed by the General health Questionnaire -GHQ-5 and levels of burnout was assessed using the ICMR-NIOH Burnout questionnaire. Multivariable logistic regression analysis was performed to identify factors associated with the risk of psychological distress. The third quartile values of the three subscales of burnout viz EE, DP and PA were used to identify burnout profiles of the healthcare workers. RESULTS: Overall, 52.9% of the participants had the risk of psychological distress that needed further evaluation. Risk of psychological distress was significantly associated with longer hours of work (≥ 8 hours a day) (AOR = 2.38, 95% CI(1.66-3.41), income≥20000(AOR = 1.74, 95% CI, (1.16-2.6); screening of COVID-19 patients (AOR = 1.63 95% CI (1.09-2.46), contact tracing (AOR = 2.05, 95% CI (1.1-3.81), High Emotional exhaustion score (EE ≥16) (AOR = 4.41 95% CI (3.14-6.28) and High Depersonalisation score (DP≥7) (AOR = 1.79, 95% CI (1.28-2.51)). About 4.7% of the HCWs were overextended (EE>18); 6.5% were disengaged (DP>8) and 9.7% HCWs were showing signs of burnout (high on all three dimensions). CONCLUSION: The study has identified key factors that could have been likely triggers for psychological distress among healthcare workers who were engaged in management of COVID cases in India. The study also demonstrates the use of GHQ-5 and ICMR-NIOH Burnout questionnaire as important tools to identify persons at risk of psychological distress and occurrence of burnout symptoms respectively. The findings provide useful guide to planning interventions to mitigate mental health problems among HCW in future epidemic/pandemic scenarios in the country.

Chitosan nanoparticles for controlled delivery of brimonidine tartrate to the ocular membrane.

Various efforts have been made to improve the bioavailability and to prolong the residence time of eye drops. Drug loaded polymeric nanoparticles offer several favorable biological properties. Thus, brimonidine tartrate (BT) loaded chitosan (CS) nanoparticles were prepared by inducing the ionic gelation upon addition of sodium tripolyphosphate (TPP). Nanoparticles were characterized by TEM, SEM, particle size, polydispersity index (PI), DSC, IR, entrapment efficiency which gave an insight of physicochemical interaction that influenced the CS nanoparticle formation and entrapment of BT. In vitro release of BT nanoparticle showed sustained release over the period of 4 h in saline phosphate buffer pH 7.4. Both placebo and BT loaded nanoparticles had a mean particle size range of about 270-370 nm with PI less than 0.5. DSC studies demonstrated structural interactions between BT, TPP and CS matrix. Entrapment efficiency of the CS nanoparticles ranged from 36-49% depending on the CS:TPP weight ratio. In vivo studies confirmed a significant sustained effect of BT nanoparticles compared to conventional eye drops. These results suggest that BT loaded CS nanoparticles could help to reduce dosage frequency by sustained drug release in the treatment of glaucoma.

Ileosigmoid knot - A Surgeon's Nightmare.

Ileosigmoid knot, also known as compound volvulus, is an unusual and a rare cause of intestinal obstruction. We are reporting a case of ileosigmoid knot in a 30-year-old male, who presented with lower abdominal pain. On examination, there was tenderness in the suprapubic area and later, the patient developed features of peritonitis. Exploratory laparotomy revealed a large volume of haemorrhagic fluid with gangrenous sigmoid colon, distended and gangrenous ileum twisted round the base of the sigmoid loop. Gangrenous portion of the ileum and sigmoid colon was resected and end to end anastomosis with Hartmann's procedure was done.

Spectrum of perforation peritonitis.

INTRODUCTION: Perforation peritonitis mostly results from the perforation of a diseased viscus. Other causes of perforation include abdominal trauma, ingestion of sharp foreign body and iatrogenic perforation. The diagnosis is mainly based on clinical grounds. Plain abdominal X-rays (erect) may reveal dilated and oedematous intestines with pneumoperitoneum. Ultrasound and CT scan may diagnose up to 72% and 82% of perforation respectively. The present study was carried out to study various etiological factors, modes of clinical presentation, morbidity and mortality patterns of perforation peritonitis presented in the RIMS hospital, Imphal, India. MATERIAL AND METHODS: The study was conducted from September 2010 to August 2012 on 490 cases of perforation peritonitis admitted and treated in the Department of Surgery. Initial diagnosis was made on the basis of detailed history, clinical examination and presence of pneumoperitoneum on erect abdominal X-ray. RESULTS: A total of 490 patients of perforation peritonitis were included in the study, with mean age of 48.28 years. 54.29% patients were below 50 years and 45.71% patients were above 50 years. There were 54.29% male patients and 45.71% female patients. Only 30% patients presented within 24 hours of onset of symptoms, 31.43% patients presented between 24 to 72 hours and 38.57% patients presented 72 hours after the onset of symptoms. Mean duration of presentation was 54.7 hours. Overall 469 patients were treated surgically and 21 patients were managed conservatively. Overall morbidity and mortality recorded in this study were 52.24% and 10% respectively.

Ligand-based Pharmacophore Modeling; Atom-based 3D-QSAR Analysis and Molecular Docking Studies of Phosphoinositide-Dependent Kinase-1 Inhibitors.

Phosphoinositide-dependent kinase-1 plays a vital role in the PI3-kinase signaling pathway that regulates gene expression, cell cycle growth and proliferation. The common human cancers include lung, breast, blood and prostate possess over stimulation of the phosphoinositide-dependent kinase-1 signaling and making phosphoinositide-dependent kinase-1 an interesting therapeutic target in oncology. A ligand-based pharmacophore and atom-based 3D-QSAR studies were carried out on a set of 82 inhibitors of PDK1. A six point pharmacophore with two hydrogen bond acceptors (A), three hydrogen bond donors (D) and one hydrophobic group (H) was obtained. The pharmacophore hypothesis yielded a 3D-QSAR model with good partial least square statistics results. The training set correlation is characterized by partial least square factors (R(2) = 0.9557, SD = 0.2334, F = 215.5, P = 1.407e-32). The test set correlation is characterized by partial least square factors (Q(2) ext = 0.7510, RMSE = 0.5225, Pearson-R =0.8676). The external validation indicated that our QSAR model possess high predictive power with good value of 0.99 and value of 0.88. The docking results show the binding orientations of these inhibitors at active site amino acid residues (Ala162, Thr222, Glu209 and Glu166) of phosphoinositide-dependent kinase-1 protein. The binding free energy interactions of protein-ligand complex have been calculated, which plays an important role in molecular recognition and drug design approach.

Comparative evaluation of intramuscular buprenorphine, pentazocine and nefopam in post-operative pain relief.

In immediate postoperative period, many injectable analgesics are being used with diverse side effects. The present study was conducted on 75 patients of both sexes, who underwent various types of surgery, to evaluate pain relief in immediate postoperative period. The drugs used for producing postoperative analgesia are buprenorphine, pentazocine and nefopam, all by intramuscular route.

Bilateral scrotal urinary extravasation in a transplant recipient mimicking acute filariasis.

Urinary leak following renal transplantation is less common nowadays with better techniques. Bilateral scrotal extravasation following urinary leak is extremely rare. A history of filariasis in an endemic zone could mask and delay the diagnosis. One such case is described wherein ureteroneocystostomy was revised and the patient could be successfully managed following urinary leak.