In silico designing and characterization of outer membrane protein K (OmpK) from Vibrio anguillarum and its expression in Nicotiana tabacum for the development of a plant-based vaccine against fish vibriosis.

Vibriosis is caused by Vibrio anguillarum in various species of aquaculture. A novel, secure, and stable vaccine is needed to eradicate vibriosis. Here, for reverse vaccinology and plant-based expression, the outer membrane protein K (OmpK) of V. anguillarum was chosen due to its conserved nature in all Vibrio species. OmpK, an ideal vaccine candidate against vibriosis, demonstrated immunogenic, non-allergic, and non-toxic behavior by using various bioinformatics tools. Docking showed the interaction of the OmpK model with TLR-5. In comparison to costly platforms, plants can be used as alternative and economic bio-factories to produce vaccine antigens. We expressed OmpK antigen in Nicotiana tabacum using Agrobacterium-mediated transformation. The expression vector was constructed using Gateway® cloning. Transgene integration was verified by polymerase chain reaction (PCR), and the copy number via qRT-PCR, which showed two copies of transgenes. Western blotting detected monomeric form of OmpK protein. The total soluble protein (TSP) fraction of OmpK was equivalent to 0.38% as detected by ELISA. Mice and fish were immunized with plant-derived OmpK antigen, which showed a significantly high level of anti-OmpK antibodies. The present study is the first report of OmpK antigen expression in higher plants for the potential use as vaccine in aquaculture against vibriosis, which could provide protection against multiple Vibrio species due to the conserved nature OmpK antigen.

Characterization and Evaluation of Silk Sericin-Based Hydrogel: A Promising Biomaterial for Efficient Healing of Acute Wounds.

The present study was aimed to prepare the potent silk sericin-based hydrogels in combination with plant extracts (curcumin and banana peel powder) and silver nanoparticles (AgNPs) to accelerate the acute wound healing process. Experimental excision wounds were created in mice by biopsy puncture, and the wound healing potential of silk sericin (2%)-based hydrogel and its combinations with curcumin (2%), banana peel powder (2%), and AgNPs (2%) was estimated by calculating the percent wound contraction, healing time, histology of skin tissues, and different biochemical tests. The results showed that the mice treated with sericin-based hydrogels showed significantly (P < 0.001) high percent wound contraction as compared to negative control, and wounds were healed in 11 days. The histological evaluation also showed that wounds covered with hydrogels were healed more than the uncovered wounds. Furthermore, the results of biochemical tests revealed that the treatment groups showed a significant (P < 0.001) decrease in the serum level of pro-inflammatory cytokines (IL-6). A significant (P < 0.001) increase in anti-inflammatory cytokines (IL-10) and anti-oxidant enzymes was observed in treatment groups. The highest wound healing potential was observed by sericin-based hydrogel containing banana peel powder, leaving behind the commercially available ointment polyfax (positive control). It can be concluded that the silk sericin-based hydrogels in combination with plant extract and AgNPs can be used as natural biomaterials in wound dressing for the rapid healing of acute wounds.

Effect of amino acid fortified mulberry leaves on economic and biological traits of Bombyx mori L.

The demand for silk has been increasing day by day but the average silk production is not enough to meet its demand. In this study, we investigated the effect of amino acid supplemented mulberry feed on the biological and commercial traits of Bombyx mori L. (Lepidoptera; Bombycidae). The silkworm larvae at 5th instar stage were taken and fed with fresh and healthy mulberry leaves coated with Alanine, Glycine and Serine in fourteen different combinations. Results of the current study revealed that the average weight of silkworm larvae and the % ratio of silk gland to body weight on day 7 was significantly (P˂0.05) higher in the group fed with amino acid fortified leaves as compared to the control. The commercial traits of larvae fed with amino acid fortified leaves also improved significantly. The larvae fed with Alanine (1 %) treated mulberry leaves showed the maximum cocoon weight, cocoon length, cocoon width, cocoon shell ratio and fibroin content as compared to the control group. It is evident from the results that the amino acid (particularly alanine) coated mulberry leaves have a positive effect on the commercial and biological traits of Bombyx mori (L.).

Biomolecules based hydrogels and their potential biomedical applications: A comprehensive review.

The need for biocompatible drug carriers has been significantly increased from the past few years. Researchers show great interest in the development of more versatile and sophisticated biomaterials based drug carriers. Hydrogels are beneficial drug carriers and easily release the controlled amount of drug at target site due to its tunable structure. The hydrogels made-up of potent biological macromolecules including collagen, gelatin, fibrin, elastin, fibroin, chitosan, starch, alginate, agarose and carrageenan have been proven as versatile biomaterials. These are three-dimensional polymeric networks, synthesized by crosslinking of hydrophilic polymers. The biological macromolecules based hydrogels containing therapeutic substances are used in a wide range of biomedical applications including wound healing, tissue engineering, cosmetics and contact lenses. However, many aspects related to hydrogels such as the mechanism of cross-linking and molecular entanglement are not clear. So, there is a need to do more research and exploration toward the extensive and cost-effective use of hydrogels. The present review article elaborately discusses the biomolecules based hydrogels and their possible biomedical applications in different fields.

Association of Guillain-Barre syndrome with COVID-19 infection: An updated systematic review.

OBJECTIVE: The systematic review aimed to determine demographic characteristics, clinical features, lab evaluation, management and complications of the studies focusing on Guillain-Barre syndrome (GBS) as a sequele of novel coronavirus (COVID-19) infection. METHODS: After protocol registration, PubMed, Web of Science and Cumulative Index to Nursing & Allied Health Literature (CINHAL) databases were searched for relevant articles using MeSH key-words and imported into referencing/review softwares. The data, regarding demographic and clinical characteristics, diagnostic workup and management, was analyzed in International Business Machines (IBM) Statistics SPSS 21. Many statistical tests, such as t-test and the Mann-Whitney U test, were used. P < 0.05 was considered significant. RESULTS: We identified 64 relevant articles. The mean age of the patients was 56 ± 16 years; the majority were males (64.9%). Among the neurological findings, paresthesia was the most typical symptom (48.9%). Most of the patients had been diagnosed by reverse transcriptase-polymerase chain reaction (RT-PCR) (69.2%). Two-third of the patients received immunoglobulins (IVIg) (77.7%). Although functions recovered in most patients, there were four patients with facial diplegia during follow-up (4.26%). Acute inflammatory demyelinating polyneuropathy (AIDP) was more likely to be associated with paresis of the lower extremity (p < 0.05) and higher levels of glucose on cerebrospinal fluid (CSF) analysis (p < 0.05). These patients were more likely to receive IVIg (p < 0.05) and develop respiratory insufficiency, subsequently (p < 0.05). CONCLUSIONS: GBS is being recognized as one of the many presentations of the COVID-19 infection. Although the common form is AIDP that might lead to complications, other variants are possible as well, and more studies are needed to focus on those subvariants.

Scientific Validation of Ethnomedicinal Use of Ipomoea batatas L. Lam. as Aphrodisiac and Gonadoprotective Agent against Bisphenol A Induced Testicular Toxicity in Male Sprague Dawley Rats.

Sweet potato (Ipomoea batatas L. Lam.), known as "Shakarqandi" in Pakistan, is an imperative root vegetable with large size, traditionally used as aphrodisiac, antiprostatic, anti-inflammatory, antidiabetic, cardiotonic, and anticancer agent. Present study was conducted to gauge aphrodisiac potential of Ipomoea batatas ethyl acetate (IPT-EA, IPA-EA) and methanol (IPT-M, IPA-M) extracts from tuber and aerial part, respectively, via behavioral and biochemical tests and their possible protective role in BPA-induced gonadotoxicity at the dose 300 mg/kg in male Sprague Dawley rats. Phytochemical analysis was done qualitatively and quantitatively through total phenolic and flavonoid content (TPC and TFC) and high performance liquid chromatographic (HPLC-DAD) fingerprinting while antioxidant profiling used multimode in vitro assays. To calculate sexual excitement mount latency, intromission latency, mount frequency, intromission frequency, ejaculatory latency, and postejaculatory interval were examined while for biochemical ratification semen characteristics, levels of testosterone, follicle stimulating hormone (FSH), luteinizing hormone (LH), and estradiol were measured. Gonadoprotective ability was assessed through comet assay and histomorphological examination of testes. Qualitative analysis ensured the presence of phenols, flavonoids, tannins, anthocyanin, saponins, coumarins, terpenoids, and betacyanin. Quantitatively maximal TPC (304.32±7.20 µg GAE/mg dry extract) and TFC (214.77±4.09 µg QE/mg DE) were estimated in IPA-EA extract. IPT-EA yielded maximum rutin (7.3±0.12) and myricetin (2.7±0.14 µg/mg DE) while IPA-EA and IPA-M yielded maximum caffeic acid (4.05±0.22 and 1.92±0.17 µg/mg DE, respectively) in HPLC-DAD analysis. Extracts enhanced sexual excitement, improved semen quality, levels of testosterone, FSH, LH, and estradiol, and successfully attenuated toxic effects of BPA. Levels of endogenous antioxidant enzymes (CAT, SOD, POD, and GSH) were restored and NO abundance was minimized. Significant stimulation in sexual behavior, amelioration of toxicity symptoms, elevated spermatic production, raised viability, vitalized levels of gonadal hormones, maintained endogenous enzymes, genoprotection, and reformed testicular histology endorsed I. batatas as a better aphrodisiac alternative and gonadoprotective agent.