Designing a next generation multi-epitope based peptide vaccine candidate against SARS-CoV-2 using computational approaches.

COVID-19 caused by SARS-CoV-2 was declared a global pandemic by WHO (World Health Organization) in March, 2020. Within 6 months, nearly 750,000 deaths are claimed by COVID-19 across the globe. This called for immediate social, scientific, technological, public and community interventions. Considering the severity of infection and the associated mortalities, global efforts are underway to develop preventive measures against SARS-CoV-2. Among the SARS-CoV-2 target proteins, Spike (S) glycoprotein (a.k.a S Protein) is the most studied target known to trigger strong host immune response. A detailed analysis of S protein-based epitopes enabled us to design a novel B-cell-derived T-cell Multi-epitope-based peptide (MEBP) vaccine candidate. This involved a systematic and comprehensive computational protocol consisting of prediction of dual-purpose epitopes and designing an MEBP vaccine construct. This was followed by 3D structure validation, MEBP complex interaction studies, in silico cloning and vaccine dose-based immune response simulation to evaluate the immunogenic potency of the vaccine construct. The dual-purpose epitope prediction protocol was designed such that the same epitope elicits both humoral and cellular immune response unlike the earlier designs. Further, the epitopes predicted were screened against stringent criteria to ensure selection of a potent candidate with maximum antigen coverage and best immune response. The vaccine dose-based immune response simulation studies revealed a rapid antigen clearance through antibody generation and elevated levels of cell-mediated immunity during repeated exposure of the vaccine. The favourable results of the analysis strongly indicate that the vaccine construct is indeed a potent vaccine candidate and ready to proceed to the next steps of experimental validation and efficacy studies. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s13205-020-02574-x.

Ameliorative effect of chitosan-conjugated 17α-methyltestosterone on testicular development in Clarias batrachus.

Chitosan nanoparticles conjugated with 17α-methyltestosterone (CS + MT) were used for studying their effect on the testicular development of Clarias batrachus during different reproductive phases. The size of chitosan nanoparticles was 127.2 nm and the nano-conjugated 17α methyltestosterone (17α-MT) was 196.1 nm (20 mg/100 ml of chitosan). Single injections of CS + MT at different doses such as 0.01, 0.1 and 0.5 µg/g body weight were administered to adults during the pre-spawning, spawning and post-spawning phase. Nano-conjugated steroid was effective at the lower dose; showing an increase in the Gonadosomatic Index (GSI) and 11-ketotestosterone level compared to the control group. Histological observations confirmed the dose-dependent advancement in spermatogenesis. These findings indicate the possibility of using CS + MT for enhancing gonadal maturity of C. batrachus.

Expression analysis of Sox9 genes during annual reproductive cycles in gonads and after nanodelivery of LHRH in Clarias batrachus.

Transcription factor Sox9 plays a crucial role in determining the fate of several cell types and is a primary factor in regulation of gonadal development. Present study reports full-length cDNA sequence of Sox9a gene and partial coding sequence (cds) of Sox9b (two duplicate orthologs of Sox9 gene) from Clarias batrachus. The coding region of Sox9a gene encoded a peptide of 460 amino acids. The partial cds of Sox9b with the length of 558bp was amplified that codes for 186 amino acids. Quantitative Real-time PCR (qRT-PCR) analysis revealed that Sox9a and Sox9b mRNA expression was significantly higher in gonads and brain tissues. Furthermore Sox9a and Sox9b mRNA expression levels were high during preparatory and pre-spawning phases and decreased gradually with onset of spawning and post-spawning phases of reproductive cycles in gonads. Chitosan nanoconjugated sLHRH (CsLHRH) of particle size 133.0nm and zeta potential of 34.3mV were synthesized and evaluated against naked sLHRH (salmon luteinizing hormone-releasing hormone). The entrapment efficiency of CsLHRH was 63%. CsLHRH nanoparticles increased the expression level of Sox9 transcripts in gonads and steroid hormonal levels in blood of male and female. Thus, our findings clearly indicate that Sox9 genes play essential role during seasonal variation of gonads. Besides, the current study reports that sustained release delivery-system will be helpful for proper gonadal development of fish. To the best of our knowledge, till date no study has been reported on nanodelivery of sLHRH and their effect on reproductive gene expression in fish.

Maximum aerobic capacity of underground coal miners in India.

Miners fitness test was assessed in terms of determination of maximum aerobic capacity by an indirect method following a standard step test protocol before going down to mine by taking into consideration of heart rates (Telemetric recording) and oxygen consumption of the subjects (Oxylog-II) during exercise at different working rates. Maximal heart rate was derived as 220-age. Coal miners reported a maximum aerobic capacity within a range of 35-38.3 mL/kg/min. It also revealed that oldest miners (50-59 yrs) had a lowest maximal oxygen uptake (34.2 ± 3.38 mL/kg/min) compared to (42.4 ± 2.03 mL/kg/min) compared to (42.4 ± 2.03 mL/kg/min) the youngest group (20-29 yrs). It was found to be negatively correlated with age (r = -0.55 and -0.33 for younger and older groups respectively) and directly associated with the body weight of the subjects (r = 0.57 - 0.68, P ≤ 0.001). Carriers showed maximum cardio respiratory capacity compared to other miners. Indian miners VO(2 max) was found to be lower both compared to their abroad mining counterparts and various other non-mining occupational working groups in India.

A comparison of cardiac strain among drillers of two different age groups in underground manual coal mines in India.

Cardiac strain was evaluated in terms of working heart rate (WHR), relative cardiac cost (RCC), net cardiac cost (NCC) and other recovery indices among six younger (mean age 34.2 +/- 2.7 yr) and sixteen older (mean age 48.9 +/- 5.4 yr) drillers working in a manual underground coal mine over two spells of work. The mean WHR was within the range of 117-132 beats / min with corresponding mean relative cardiac cost between 44-48% of heart rate reserve for the younger group and 53-55% for their older counterparts. The mean NCC was above 50 beats/min for both age groups. It was seen that the workers surpassed the recommended limits of cardiac strain indices. The intensity of workload indicates the job to be "heavy" to "extremely heavy" in accordance with the heaviness scales based on WHR, NCC and recovery heart rates. Heat stress prevailing in the workplace in terms of effective temperature (ET) and wet bulb globe temperature (WBGT), was above the recommended limits as per the guidelines proposed by WHO and ACGIH. High physiological demands of the job which requires predominate static muscular exertions coupled with high heat stress were found to hinder the recovery process and may prove deleterious particularly for the older workers. Therefore, in the present context, the need of ergonomic interventions for job organization and quick reparation of environmental condition are strongly indicted.

A comparison of physiological strain of carriers in underground manual coal mines in India.

Thirty-nine healthy carriers (23-57 years of age) were investigated in underground manual coal mines in West Bengal, India during two different work spells of a single work shift. We compared physiological strain of workers <40 and > or =40 years of age. For both groups, mean heart rate was 124-133 beats/min, with a mean corresponding relative cardiac cost of 50-66%. Maximum aerobic capacities were estimated indirectly, following a standard step test protocol. Average oxygen consumption was 1.07-1.1 l/min, with an energy expenditure of 5.35-5.5 kcal/min among both age groups. Acceptable levels of physiological strain were well encroached, and older workers faced the maximum burden. The tasks studied were heavy to very heavy in nature. The weight of load carriage at a spontaneously chosen speed and the prevailing environmental conditions merit serious attention. There is extreme need of ergonomic interventions in reducing the postural load and musculoskeletal discomforts in this population.

A comparative study of physiological strain of underground coal miners in India.

Ninety-eight healthy underground coal miners aged between 23-58 years were studied during their activity period. Physiological strain of different category of miners in terms of heart rate was monitored continuously with heart rate monitor that revealed the tasks as heavy to very heavy for them. Oxygen consumption was measured directly by using oxylog-2 machine that corresponded to metabolic costs for different activities ranging from 4.96 kcal/min to 5.47 kcal/min. The mean relative aerobic strain varied from 47.4%-56.8%--depicting acceptable level of physical strain was well encroached by the miners who irrespective of ages and categories showed poor recovery responses. This entails that miners are exerting themselves beyond their capacities where inevitably older workforce face the maximum burden.

A study of the workload of underground trammers in the Ranigang coal field area of West Bengal, India.

Thirty healthy trammers were examined in underground haulage mines during their normal activity. Physiological strain in terms of heart rate (HR) varied between 101.6 and 104.7 beats/min with mean net cardiac cost of 33.06 and 34.06 beats/min for younger and older groups respectively. The average relative cardiac cost was lower for younger subjects than for older ones. Oxygen consumption (VO2) was measured with an Oxylog-II machine (UK); subsequently values were estimated. Maximum aerobic capacity was estimated with an indirect method following a standard step test protocol. The responses revealed that the average VO2 during the activity was 0.75 and 0.8 L/min, which corresponded to energy expenditure (EE) of 3.8 and 3.97 Kcal/min for younger and older subjects respectively. The workload in terms of HR and EE was moderate, whereas the aerobic strain experienced by aged workers was above the acceptable level.

Cardiovascular load assessment of coal mine shovelers in west Bengal, India: a comparison between middle age groups.

Cardiovascular load of underground shovelers was assessed in terms of heart rate changes at work in haulage coalmines. Twenty underground shovelers of two different age groups (those of age 31-39 and those of age 40-49) served as subjects. Working heart rate was significantly higher (p < 0.01) in the older group (138.2 beats/min) than in the younger group (130.4 beats/min). Though the net cardiac cost did not differ significantly between the groups (64.8 beats/min and 69.4 beats/min in younger and older groups, respectively), difference in the relative cardiac cost (54.17% and 64.86% in younger and older groups, respectively) proved to be highly significant (p < 0.001). The heart rate up to the third minute of recovery showed significant differences between the groups (p < 0.01). While analysis of physiological responses indicated that the task was very strenuous irrespective of age, cardiac strain was found to be more excessive in the older subjects if continued. The arduous nature of the working environment in the coalfaces was also reported.