Immunoinformatic exploration of a multi-epitope-based peptide vaccine candidate targeting emerging variants of SARS-CoV-2.

Many countries around the world are facing severe challenges due to the recently emerging variants of SARS-CoV-2. Over the last few months, scientists have been developing treatments, drugs, and vaccines to subdue the virus and prevent its transmission. In this context, a peptide-based vaccine construct containing pathogenic proteins of the virus known to elicit an immune response was constructed. An analysis of the spike protein-based epitopes allowed us to design an "epitope-based subunit vaccine" against coronavirus using the approaches of "reverse vaccinology" and "immunoinformatics." Computational experimentation and a systematic, comprehensive protocol were followed with an aim to develop and design a multi-epitope-based peptide (MEBP) vaccine candidate. Our study attempted to predict an MEBP vaccine by introducing mutations of SARS-CoV-2 (Delta, Lambda, Iota, Omicron, and Kappa) in Spike glycoprotein and predicting dual-purpose epitopes (B-cell and T-cell). This was followed by screening the selected epitopes based on antigenicity, allergenicity, and population coverage and constructing them into a vaccine by using linkers and adjuvants. The vaccine construct was analyzed for its physicochemical properties and secondary structure prediction, and a 3D structure was built, refined, and validated. Furthermore, the peptide-protein interaction of the vaccine construct with Toll-like receptor (TLR) molecules was performed. Immune profiling was performed to check the immune response. Codon optimization of the vaccine construct was performed to obtain the GC content before cloning it into the E. coli genome, facilitating its progression it into a vector. Finally, an in-silico simulation of the vaccine-protein complex was performed to comprehend its stability and conformational behavior.

Synthesis of molnupiravir (MK-4482, EIDD-2801): a promising oral drug for the treatment of COVID-19 starting from cytidine.

The present work describes the synthesis of molnupiravir by employing commercially available inexpensive materials in two steps with an overall yield of 85.7%. The synthetic methodology starts with an eco-friendly starting material, that is, cytidine and establishes an alternative way to avoid costly enzyme mediated reactions. This synthetic strategy involves a selective acylation of cytidine as the first key step followed by the second step, that is, hydroxamination reaction. The major advantage of this protocol is that it is completely free of protection and deprotection reactions. Chemoselective acylation of cytidine's primary alcohol was achieved using isobutyryl chloride, Et3N, and DMF solvent (89.3% yield). The aqueous phase transformation was achieved for the hydroxamination reaction with a 96% yield.

High efficient somatic embryogenesis development from leaf cultures of Citrullus colocynthis (L.) Schrad for generating true type clones.

We report an efficient somatic embryogenesis and plant regeneration system using leaf cultures of Citrullus colocynthis (L.) and assessed the effect of plant growth regulators on the regeneration process. Initially leaf explants were cultured on Murashige and Skoog medium supplemented with different concentrations of auxins viz., 2,4-dichlorophenoxyacetic acid, 1-naphthaleneacetic acid, gibberellic acid alone and along with combination of 6-benzylaminopurine. The different forms of calli such as compact, white friable, creamy friable, brownish nodular, green globular and green calli were induced from the leaf explants on MS medium containing different concentrations of auxins and gibberellins. Subsequently initial callus was subcultured at 1.5 mg L(-1) BAP + 1.0 mg L(-1) 2,4-D which resulted in 25 % somatic embryos from 85 % nodular embryogenic nodular callus that is highest percentage. Similarly the lowest percentage of somatic embryos was recorded at 2.5 mg L(-1) BAP + 0.5 mg L(-1) NAA from 55 % embryogenic globular callus i.e., 16 %. High frequency of embryo development takes place at intermittent light when compared with continuous light in the individual subcultures. The cotyledonary embryos were developed into complete platelets on MS medium. In vitro regenerated plantlets were washed to remove the traces of agar and then transferred to sterile vermiculite and sand (2:1) containing pot.

Synthesis, Characterization, Fluorescence, Photocatalytic and Antibacterial Activity of CdS Nanoparticles Using Schiff Base.

Cadmium sulfide nanoparticles (CdS NPs) were successfully prepared using sonochemical method by employing Schiff-base, (2-[(4-methoxy-phenylimino)-methyl]-4-nitro phenol) as a complexing agent. Here, SB is used as a ligand to control the morphology of NPs. XRD patterns and TEM images show that the synthesized CdS NPs have cubic structures with a diameter of about 2-10 nm. The formation of CdS NPs and their optical, structure, thermal and morphologies were studied by means of UV-vis DRS, fluorescence, FTIR, zeta potential, XRD, SEM and TEM. The interactions between CdS NPs and SB were investigated in an aqueous solution using fluorescence spectroscopy. The fluorescence quenching studies suggest that SB quenches the fluorescence of CdS NPs effectively. The degradation kinetics of methyl red (MR) by the photocatalyst was followed by Langmuir-Hinshelwood model. The results revealed that photocatalytic degradation of MR by SB capped CdS NPs could be considered as a practical and reliable technique for the removal of environmental pollutants. The antibacterial activity of samples was evaluated against E. coli, S. aureus and P. aeruginosa and the results were compared. SB and SB capped CdS NPs could be a potential antibacterial compounds after further investigation.

Effect of amitriptyline on adrenergic receptor number and second messenger function in rat brain.

Radioligand binding studies were done to investigate the effect of chronic administration of Amitriptyline on alpha1-adrenoceptor (alpha1-AR) receptor mediated response to inositol triphosphate (IP3) in rat brain. Our studies revealed a significant decrease in the densities of alpha1-ARs in cortex and cerebellum of rat brain after chronic administration of Amitriptyline (10 mg kg-1 b.wt.). However, there was no significant change in the affinity of [3H]prazosin to alpha1-ARs. Displacement studies showed that Amitriptyline has higher affinity for alpha1-AR with a Ki value of 182+/-16 nM. Significant change was observed in basal IP3 activity in cortex and cerebellum after Amitriptyline exposure. In cortex and cerebellum of experimental rats the NE (Norepinephrine) stimulated IP3 activity was significantly decreased (1460+/-102 DPM/g tissue; p<0.0001; 1188+/-112 DPM/g tissue; p<0.0001), when compared to NE stimulated IP3 activity (4152+/-286 and 3952+/-245 DPM/g tissue, respectively) in control rats. The decrease in NE stimulated IP3 activity in both regions may be due to the significant downregulation of alpha1-ARs in cortex after Amitriptyline exposure as these sites are positively coupled to IP3. The observed significant decrease in alphal-ARs with concomitant decrease in NE stimulated IP3 activity, after Amitriptyline treatment, suggests that Amitriptyline which has high affinity for these sites, acts by modulating the alpha1-AR receptor mediated response in brain.

Differential modulation of α-1 adrenoceptor subtypes by antidepressants in the rat brain.

The aim of the present study was to examine the effect of chronic antidepressants treatment on the density of α1-adrenoceptor (AR) subtypes in rat brain. Density of total α1 and α(1A)- and α(1Β)-ARs was measured in cortex and cerebellum of rats treated with amitriptyline (AMI), desipramine (DMI) and fluoxetine (FLX), (10 mg/kg body wt), for 30 days, using [³H]prazosin in presence and absence of WB-4101. The density of cortical total α1-ARs was significantly decreased with AMI (54%) and DMI (25%) treatment, without altering the affinity of the receptor. Fluoxetine did not alter the density of cortical α1-ARs. The density of cortical α(1A)-ARs was also significantly decreased with AMI (85%) and DMI (50%) treatment, without affecting the affinity. The density of cerebellar total α1-ARs was significantly decreased with AMI (37%), DMI (50%) and FLX (70%) treatment, without affecting the affinity for [³H]prazosin. The density of α(1A)-ARs was significantly decreased with AMI (67%), DMI (59%) and FLX (92%) treatment. α(1B)-AR density was decreased only with FLX (47%) and DMI (47%) treatment. Correspondingly the basal IP3 and NE (10 µM) stimulated IP3 levels were significantly decreased in AMI (47%), DMI (22%) and FLX (48%) treated rat cortex. The results suggest that chronic antidepressant (AD) treatment down-regulates the cortical and cerebellar total α1-ARs in rat brain. However, α(1A) subtype is predominantly down-regulated by AMI and DMI, where as FLX affects cerebellar α(1A)-ARs. The region-specific and subtype specific down-regulation of α1-ARs density, which occurs after prolonged AD treatment, may underline the therapeutic mechanism of action.

In vivo evaluation of a closed loop monitoring strategy for induced paralysis.

OBJECTIVE: Reliable closed loop infusion systems for regulating paralysis level can be a great convenience to the anesthesiologists in automating their task. This paper describes the in vivo performance evaluation of a self-tuning controller that is designed to accommodate large variations in patient drug sensitivity, drug action delays and environmental interfering noise. METHODS: The infusion system was evaluated in six adult mongrel dogs. Following the manual induction of paralysis by an anesthesiologist, the controller regulated the infusion of vecuronium to maintain a desired level of paralysis. The integrated EMG response of the hypothenar muscle to a train-of-four stimulation of the ulnar nerve quantified the depth of paralysis. The controller's robustness was tested by contaminating the sensed twitch signal with electrocautery noise and electrode disconnection. RESULTS: The controller reached the initial level of paralysis of 100% in about 4.0 minutes and arrived at the desired level of 90% with an overshoot of 6.38% (+/-6.82). It maintained the desired level of paralysis with a 2.04% (+/-1.20) mean offset at 90% and 0.4% (+/-0.5) mean offset at 80% steady state level, respectively. The mean infusion rate to sustain 90% and 80% paralysis were 2.70 (+/-2.05) and 2.15 (+/-2.57) ((mg/kg)/min), respectively. CONCLUSIONS: The system adapted to a large variation in the sample subject drug sensitivity. It remained stable despite large amplitude disturbances and maintained the paralysis at the desired level following the removal of the disturbances.

A controller for automatic regulation of induced paralysis during surgery.

This paper presents a strategy for automatic control of induced paralysis using vecuronium bromide during surgery. The controller is self-tuning and adapts to inter-patient and intra-patient response variations while optimizing the output variance and infusion rate. In particular, the controller is capable of accommodating the variations of pure time delays in patient response. The performance of the controller is evaluated using an experimentally derived pharmacokinetic and nonlinear pharmacodynamic model of patient response. The results indicate that the controller provides robust regulation of the paralysis level with no output offset.