Cyanobacterial Proteomics: Diversity and Dynamics.

Cyanobacteria (oxygenic photoautrophs) comprise a diverse group holding significance both environmentally and for biotechnological applications. The utilization of proteomic techniques has significantly influenced investigations concerning cyanobacteria. Application of proteomics allows for large-scale analysis of protein expression and function within cyanobacterial systems. The cyanobacterial proteome exhibits tremendous functional, spatial, and temporal diversity regulated by multiple factors that continuously modify protein abundance, post-translational modifications, interactions, localization, and activity to meet the dynamic needs of these tiny blue greens. Modern mass spectrometry-based proteomics techniques enable system-wide examination of proteome complexity through global identification and high-throughput quantification of proteins. These powerful approaches have revolutionized our understanding of proteome dynamics and promise to provide novel insights into integrated cellular behavior at an unprecedented scale. In this Review, we present modern methods and cutting-edge technologies employed for unraveling the spatiotemporal diversity and dynamics of cyanobacterial proteomics with a specific focus on the methods used to analyze post-translational modifications (PTMs) and examples of dynamic changes in the cyanobacterial proteome investigated by proteomic approaches.

Pretilachlor-induced physiological, biochemical and morphological changes in Indian paddy field agroecosystem inhabited Anabaena doliolum.

Pretilachlor is a systemic, pre-emergence herbicide applied in the paddy fields to kill narrow and broadleaf weeds. The present study evaluates the toxicity of pretilachlor on the non-target diazotrophic free-living cyanobacterium Anabaena doliolum, commonly found in the paddy fields of eastern Uttar Pradesh (India) and used as a biofertilizer. A.doliolum was subjected to several doses (0, 2, 5, 7, 10, 20 and 40 µg/ml) of pretilachlor and its effects were examined in terms of alterations in cellular morphology, ultrastructure, physiology, and biochemical attributes. The treatment of pretilachlor decreased the growth, total pigment content and photosynthetic efficiency of the test organism in a dose-dependent manner. The decline in growth was observed on 20th day at 2, 5, 7, 10, 20 and 40 µg/ml of pretilachlor concentration by 4, 9, 26, 47, 71 and 92%, respectively. Furthermore, Chlorophyll a and phycocyanin levels were noticeably declined. As a result, the photosynthetic performance also registered a similar decline as measured by chlorophyll fluorescence. However, carotenoid content increased by 13%, 41% and 53% at 5, 10 and 20 µg/ml on 5th day reflecting its protective property. A marked increase in fluorescence intensity and malondialdehyde content by 2.65 and 2.45 folds at 10 and 20 µg/ml on 7th day was registered. The enzymatic antioxidants (SOD and CAT) and a concurrent increase in glutathione reductase activity were registered (1.75 and 2.11-fold at 20 and 40 µg/ml on 5th day), indicating pretilachlor mediated ROS generation. Moreover, ultrastructural studies done by SEM and TEM revealed plasma membrane and thylakoid membrane damage and fragmentation. These findings have contributed to the broader comprehension of the stress responses triggered by pretilachlor in cyanobacteria. Moreover, they can aid in the evaluation of the detrimental impact of pretilachlor on A. doliolum, given their crucial function as a nitrogen contributor in paddy fields.

Phycochemistry and bioactivity of cyanobacterial secondary metabolites.

Microbes are a huge contributor to people's health around the world since they produce a lot of beneficial secondary metabolites. Cyanobacteria are photosynthetic prokaryotic bacteria cosmopolitan in nature. Adaptability of cyanobacteria to wide spectrum of environment can be contributed to the production of various secondary metabolites which are also therapeutic in nature. As a result, they are a good option for the development of medicinal molecules. These metabolites could be interesting COVID-19 therapeutic options because the majority of these compounds have demonstrated substantial pharmacological actions, such as neurotoxicity, cytotoxicity, and antiviral activity against HCMV, HSV-1, HHV-6, and HIV-1. They have been reported to produce a single metabolite active against wide spectrum of microbes like Fischerella ambigua produces ambigols active against bacteria, fungi and protozoa. Similarly, Moorea producens produces malygomides O and P, majusculamide C and somocystinamide which are active against bacteria, fungi and tumour cells, respectively. In addition to the above, Moorea sp. produce apratoxin A and dolastatin 15 possessing anti cancerous activity but unfortunately till date only brentuximab vedotin (trade name Adcetris), a medication derived from marine peptides, for the treatment of Hodgkin lymphoma and anaplastic large cell lymphoma has been approved by FDA. However, several publications have effectively described and categorised cyanobacterial medicines based on their biological action. In present review, an effort is made to categorize cyanobacterial metabolites on the basis of their phycochemistry. The goal of this review is to categorise cyanobacterial metabolites based on their chemical functional group, which has yet to be described.

Inorganic clay nanocomposite system for improved cholinesterase inhibition and brain pharmacokinetics of donepezil.

Objective: Brain drug delivery for effective treatment of neurodegenerative disorders is limited due to the selective permeability of blood brain barrier (BBB). During the past few years, development of novel delivery system has attracted considerable attention of formulation scientists to overcome the permeability limitation caused by BBB.Significance: Based on the outcomes of this study and taking into consideration of the unique characteristics of laponite, it can be further explored to deliver many other central nervous system acting drugs.Methods: In the present study, laponite (LAP) nanocomposites were exploited for the improved brain delivery of donepezil (DZ) following encapsulation approach due to their nano-size and positive charge at pH <9.Result: The size of prepared nanocomposites was 53.7 ± 4.0 to 137.7 ± 11.0 nm. The drug was released in a sustained manner till 120 h in phosphate buffer saline (pH 7.4) and acid phthalate buffer (pH 4.0). LAPDZ formulations inhibited acetylcholinesterase approximately by 82%, significantly higher (p < 0.05) than plain DZ (30%). Swiss albino mice exhibited enhanced brain uptake of LAPDZ administered via intravenous route. Promising pharmacokinetic parameters were observed in animals treated with LAPDZ. LAPDZ formulation showed half-life (t1/2), volume of distribution (Vd) and clearance (Cl) as 5.53 ± 0.40 h-1, 0.129 ± 0.02 L, 0.015 ± 0.002 L/h, respectively. While DZ solution showed the same parameters as 1.06 ± 0.12 h-1, 0.168 ± 0.01 L, 0.106 ± 0.013 L/h, respectively. The brain uptake of LAPDZ formulation was improved with quintuplet t1/2.Conclusion: Based on the results of present study, it is proposed that the formulated nanocomposite would result in improved patient compliance with therapeutic effect at lower doses.

BIMANUAL MICROINCISION VITREOUS SURGERY FOR SEVERE PROLIFERATIVE DIABETIC RETINOPATHY: OUTCOME IN MORE THAN 300 EYES.

PURPOSE: To evaluate the visual and anatomical outcomes and safety of bimanual microincision vitreous surgery for severe proliferative diabetic retinopathy. METHODS: Retrospective review of 315 eyes of 282 patients who underwent 23-gauge or 25-gauge pars plana vitrectomy with bimanual membrane dissection for diabetic tractional detachment from January 2007 to September 2016. Minimum follow-up was 3 months, and the average duration of follow-up was 23 months (range 3-100 months; median 15 months). Outcome measures were best-corrected visual acuity, anatomical success, and postoperative complications. RESULTS: Postoperatively, 84.3% of eyes improved (>2 lines), 10.5% were stable, and 5.4% worsened (>2 lines). Comparing gauges, two-line improvement was seen in 87.4% of 23-gauge eyes compared with 79.7% of 25-gauge eyes (P = 0.029). Mean peak best-corrected visual acuity improved from 20/930 (1.67 ± 0.63) preoperatively to 20/120 (0.78 ± 0.63) postoperatively (P < 0.001). Primary reattachment was achieved in 310 eyes (98.4%) and final reattachment in 312 eyes (99%). Recurrent vitreous hemorrhage was the commonest postoperative complication (18.4%). Lower incidence of recurrent vitreous hemorrhage was seen with 25 gauge (13.5%) compared with 23 gauge (22%, P = 0.038). Epiretinal membrane formation (7.9%), intractable glaucoma (2.5%), and endophthalmitis (0.6%) were some of the other postoperative complications. CONCLUSION: Sustained visual improvement, anatomical restoration, and low complication rates were obtained in complex situations with bimanual microincision vitreous surgery in a large series. Visual outcomes were poorer in older age group, tractional retinal detachments involving macula, and eyes with extensive membranes and with silicone oil as tamponade. Both 23-gauge and 25-gauge groups were comparable in relation to visual improvement, anatomical success, and intraoperative and postoperative complications.

Nickel and arsenite-induced differential oxidative stress and antioxidant responses in two Anabaena species.

In recent years, release of chemical pollutants has increased due to anthropogenic activities. Heterocystous filamentous cyanobacteria constitute dominant paddy microflora and are excellent biofertilizers augmenting rice productivity. Cyanobacteria are frequently exposed to toxic metals, nickel and arsenic are one of the major toxicants present. We exposed two species of diazotrophic cyanobacteria Anabaena sp. PCC 7120 and Anabaena doliolum, to sub-lethal concentrations (15.0 and 9.0 µM) of Ni2+ and (17.0 and 11 mM) of arsenite (AsIII) and analyzed at different days of treatments (0, 1, 7, and 15 days) for oxidative damage and antioxidative biomarkers. Lipid peroxidation was enhanced (1.5- to 2.5-fold increase in MDA content), indicating damaging effects of Ni2+ and As(III) on membrane. Although Ni2+ and As(III), both induced oxidative stress in both species, Anabaena PCC 7120 experienced less stress than A. doliolum. This could be explained by a higher activity of antioxidant enzymes catalase (CAT), ascorbate peroxidase (APX), glutathione reductase (GR) in Anabaena PCC 7120 (4.6-, 2.0- and 1.4-fold [Ni2+ ] 3.2-, 2.5-, and 2.08-fold [As]) compared to A. doliolum (4.2-, 2.5-, and 1.3-fold [Ni2+ ] and 3.2-, 3.33-, and 1.8-fold [As]). Moreover, superoxide dismutase registered less inhibition in Anabaena sp. PCC 7120 (1.5 and 1.8) compared to A. doliolum (1.8 and 2.3) under Ni2+ and As(III) stress. In addition to, IBR revealed that As(III) imposes severe impact on both strain, however, A. doliolum suffers most. Therefore, the study demonstrates interspecies variation in survival strategy of two Anabaena species and difference in potential of two different toxicants to produce oxidative stress.

UV-B-Heavy Metal Interaction: Comparative Evaluation on Photosynthetic Properties of Three Cyanobacteria.

Gradual increase in UV-B component reaching earth surface together with heavy metal contamination appears to be a serious environmental concern. Differential sensitivity in photosynthetic characters of Anabaena doliolum, Microcystis sp., and Nostoc muscorum were observed following exposure to UV-B radiation and heavy metals (Cd and Zn) which displayed reduced photosynthesis with maximum inhibition in Anabaena. PSII was the most sensitive component of the electron transport chain depicting 84, 80, and 70 % inhibition in A. doliolum, Microcystis sp., and N. muscorum, respectively. Cadmium and UV-B-induced inhibition of DCPIP photoreduction could not reversed by artificial electron donors in all the strains. However, they substantially reversed the inhibition caused by Zn as well as Zn + UV-B interactively in N. muscorum, not observed in other two strains. Absorption spectra of all the strains showed differential decrement in chl a peak in treated than the control cells (P < 0.25, r = -0.942). Significantly decreased peaks (P < 0.05) by different states of stresses to all the three cyanobacteria were observed in emission spectra. Excitation spectra of the test strains suggest disorganization or delinking of phycobilisomes from the PSII reaction center, also attested by strong negative correlation between the treatment of stress and phycocyanin (P < 0.025, r = -0.971). The present study qualified N. muscorum as most tolerant followed by Microcystis sp., whereas A. doliolum emerged as the most sensitive one and also endorses high toxicity of Cd as compared to Zn.

Effects of UV-B and heavy metals on nitrogen and phosphorus metabolism in three cyanobacteria.

Cyanobacteria sp. (diazotrophic and planktonic) hold a major position in ecosystem, former one due to their intrinsic capability of N2-fixation and later because of mineralization of organic matter. Unfortunately, their exposure to variety of abiotic stresses is unavoidable. Comparative analysis of interactive effect of UV-B and heavy metals (Cd/Zn) on nitrogen and phosphorus metabolism of three cyanobacteria (Anabaena, Microcystis, Nostoc) revealed additive inhibition (χ(2) significant p < 0.05) of NH4(+) and PO4(3-) uptake whereas increase in nitrate uptake upon UV-B + heavy metal exposure. Glutamine synthetase and Alkaline phosphatase activity was reduced after all treatments whereas Nitrate reductase activity showed slight stimulation in UV-B and UV-B + heavy metals treatment. Combination of UV-B and metals seems more detrimental to the NH4(+), PO4(3-) uptake, GS and APA activity. A significant stimulation in NO3(-) uptake and NR activity following exposure to UV-B in all the three cyanobacteria suggests UV-B-induced structural change(s) in the enzyme/carriers. Metals seem to compete for the binding sites of the enzymes and carriers; as noticed for Anabaena and Microcystis showing change in Km while no change in the Km value of Nostoc suggests non-competitive nutrient uptake. Higher accumulation and more adverse effect on Na(+) and K(+) efflux proposes Cd as more toxic compared to Zn.

Comparative interactions of withanolides and sterols with two members of sterol glycosyltransferases from Withania somnifera.

BACKGROUND: Sterol glycosyltransferases (SGTs) are ubiquitous but one of the most diverse group of enzymes of glycosyltransferases family. Members of this family modulate physical and chemical properties of secondary plant products important for various physiological processes. The role of SGTs has been demonstrated in the biosynthesis of pharmaceutically important molecules of medicinal plants like Withania somnifera. RESULTS: Analysis suggested conserved behaviour and high similarity in active sites of WsSGTs with other plant GTs. Substrate specificity of WsSGTs were analysed through docking performance of WsSGTs with different substrates (sterols and withanolides). Best docking results of WsSGTL1 in the form of stable enzyme-substrate complex having lowest binding energies were obtained with brassicasterol, transandrosteron and WsSGTL4 with solasodine, stigmasterol and 24-methylene cholesterol. CONCLUSION: This study reveals topological characters and conserved nature of two SGTs from W. somnifera (WsSGTs) i.e. WsSGTL1 and WsSGTL4. However, besides being ubiquitous in nature and with broad substrate specificity, difference between WsSGTL1 and WsSGTL4 is briefly described by difference in stability (binding energy) of enzyme-substrate complexes through comparative docking.

WsSGTL1 gene from Withania somnifera, modulates glycosylation profile, antioxidant system and confers biotic and salt stress tolerance in transgenic tobacco.

Glycosylation of sterols, catalysed by sterol glycosyltransferases (SGTs), improves the sterol solubility, chemical stability and compartmentalization, and helps plants to adapt to environmental changes. The SGTs in medicinal plants are of particular interest for their role in the biosynthesis of pharmacologically active substances. WsSGTL1, a SGT isolated from Withania somnifera, was expressed and functionally characterized in transgenic tobacco plants. Transgenic WsSGTL1-Nt lines showed an adaptive mechanism through demonstrating late germination, stunted growth, yellowish-green leaves and enhanced antioxidant system. The reduced chlorophyll content and chlorophyll fluorescence with decreased photosynthetic parameters were observed in WsSGTL1-Nt plants. These changes could be due to the enhanced glycosylation by WsSGTL1, as no modulation in chlorophyll biogenesis-related genes was observed in transgenic lines as compared to wildtype (WT) plants. Enhanced accumulation of main sterols like, campesterol, stigmasterol and sitosterol in glycosylated form was observed in WsSGTL1-Nt plants. Apart from these, other secondary metabolites related to plant's antioxidant system along with activities of antioxidant enzymes (SOD, CAT; two to fourfold) were enhanced in WsSGTL1-Nt as compared to WT. WsSGTL1-Nt plants showed significant resistance towards Spodoptera litura (biotic stress) with up to 27 % reduced larval weight as well as salt stress (abiotic stress) with improved survival capacity of leaf discs. The present study demonstrates that higher glycosylation of sterols and enhanced antioxidant system caused by expression of WsSGTL1 gene confers specific functions in plants to adapt under different environmental challenges.

Salinity pretreatment synergies heat shock toxicity in cyanobacterium Anabaena PCC7120.

This study was undertaken to bridge the knowledge gap pertaining to cyanobacteria's response to pretreatment. The result elucidates the synergistic effect of pretreatment toxicity in cyanobacterium Anabaena PCC7120 on morphological and biochemical attributes. Chemical (salt) and physical (heat) stress-pretreated cells exhibited significant and reproducible changes in terms of growth pattern, morphology, pigments, lipid peroxidation, and antioxidant activity. Salinity pretreatment showed more than a five-fold decrease in the phycocyanin content but a six-fold and five-fold increase in carotenoid, lipid peroxidation (MDA content), and antioxidant activity (SOD and CAT) at 1 h and on 3rd day of treatment, respectively, giving the impression of stress-induced free radicals that are scavenged by antioxidants when compared to heat shock pretreatment. Furthermore, quantitative analysis of transcript (qRT-PCR) for FeSOD and MnSOD displayed a 3.6- and 1.8-fold increase in salt-pretreated (S-H) samples. The upregulation of transcript corresponding to salt pretreatment suggests a toxic role of salinity in synergizing heat shock. However, heat pretreatment suggests a protective role in mitigating salt toxicity. It could be inferred that pretreatment enhances the deleterious effect. However, it further showed that salinity (chemical stress) augments the damaging effect of heat shock (physical stress) more profoundly than physical stress on chemical stress possibly by modulating redox balance via activation of antioxidant responses. Our study reveals that upon pretreatment of heat, the negative effect of salt can be mitigated in filamentous cyanobacteria, thus providing a foundation for improved cyanobacterial tolerance to salt stress.

Promising role of Vitamin D and plant metabolites against COVID-19: Clinical trials review.

Vitamin D possesses immunomodulatory qualities and is protective against respiratory infections. Additionally, it strengthens adaptive and cellular immunity and boosts the expression of genes involved in oxidation. Experts suggested taking vitamin D supplements to avoid and treat viral infection and also COVID-19, on the other hand, since the beginning of time, the use of plants as medicines have been vital to human wellbeing. The WHO estimates that 80 % of people worldwide use plants or herbs for therapeutic purposes. Secondary metabolites from medicinal plants are thought to be useful in lowering infections from pathogenic microorganisms due to their ability to inhibit viral protein and enzyme activity by binding with them. As a result, this manuscript seeks to describe the role of vitamin D and probable plant metabolites that have antiviral activities and may be complementary to the alternative strategy against COVID-19 in a single manuscript through reviewing various case studies.

Incidence of post vitrectomy endophthalmitis in India - A multicentric study by VRSI study Group.

INTRODUCTION: The incidence of post vitrectomy endophthalmitis (PVE) is reported to be between 0.02 and 0.84%. Resterilization of single use instruments is a common practice amidst developing countries to make it more affordable to the patients by reducing the cost of the surgery and also reduce the environmental hazard. The aim of our study is to evaluate the incidence of PVE amidst existing sterilization practices of reused instruments in multiple vitreoretinal centres in India. METHODOLOGY: Centres with an endophthalmitis tracking system were invited to participate in a survey. Twenty-five centres were sent a questionnaire via email. The questionnaire included details about the institution, number of vitrectomies performed in a year, sterilization practices followed pre-operatively, intraoperatively and postoperatively, incidence of endophthalmitis and instrument reuse policies. RESULTS: A total of 29 cases of endophthalmitis were reported out of the 47,612 vitrectomies performed across various centres. The mean incidence of endophthalmitis was 0.06%. There was no difference in the rates of endophthalmitis based on various pre-operative, intraoperative or postoperative prophylactic measures. Nearly 80% of the centres change most of the instruments after every case, while the rest reused. The mean number of times a cutter was being reused until discarded was 4.7. Nearly 76% followed a performance-based protocol, and the remaining 24% had a fixed protocol for the number of times an instrument can be reused before discarding it. CONCLUSION: PVE rates are not significantly different in India despite the multiuse of single use instruments. The purpose of this paper is not to suggest an alternate protocol but to creating one in the future with these results in mind, to rationalise the use of single use instruments, make VR surgery more affordable and also have a positive impact on the carbon footprint of consumables in surgery.

Single-Cell Transcriptome Analysis Reveals the Role of Pancreatic Secretome in COVID-19 Associated Multi-organ Dysfunctions.

The SARS-CoV-2 infection affects the lungs, heart, kidney, intestine, olfactory epithelia, liver, and pancreas and brings forward multi-organ dysfunctions (MODs). However, mechanistic details of SARS-CoV-2-induced MODs are unclear. Here, we have investigated the role of pancreatic secretory proteins to mechanistically link COVID-19 with MODs using single-cell transcriptome analysis. Secretory proteins were identified using the Human Protein Atlas. Gene ontology, pathway, and disease enrichment analyses were used to highlight the role of upregulated pancreatic secretory proteins (secretome). We show that SARS-CoV-2 infection shifts the expression profile of pancreatic endocrine cells to acinar and ductal cell-specific profiles, resulting in increased expression of acinar and ductal cell-specific genes. Among all the secretory proteins, the upregulated expression of IL1B, AGT, ALB, SPP1, CRP, SERPINA1, C3, TFRC, TNFSF10, and MIF was mainly associated with disease of diverse organs. Extensive literature and experimental evidence are used to validate the association of the upregulated pancreatic secretome with the coagulation cascade, complement activation, renin-angiotensinogen system dysregulation, endothelial cell injury and thrombosis, immune system dysregulation, and fibrosis. Our finding suggests the influence of an upregulated secretome on multi-organ systems such as nervous, cardiovascular, immune, digestive, and urogenital systems. Our study provides evidence that an upregulated pancreatic secretome is a possible cause of SARS-CoV-2-induced MODs. This finding may have a significant impact on the clinical setting regarding the prevention of SARS-CoV-2-induced MODs.

LONGITUDINAL SWEPT SOURCE OPTICAL COHERENCE TOMOGRAPHY ANGIOGRAPHY OF EXTRAFOVEAL POLYPS AFTER COMBINATION THERAPY.

PURPOSE: To study the efficacy of swept source optical coherence tomography angiography (SSOCTA) to longitudinally follow-up patients with extrafoveal polyps post-laser photocoagulation and anti-vascular endothelial growth factor injection. METHODS: Observational case series. Four patients diagnosed as polypoidal choroidal vasculopathy with extrafoveal polyps on multimodal imaging were followed up serially on SSOCT, en face and cross-sectional SSOCTA at a month and then 3 monthly for a year. Indocyanine green angiography was repeated at 4 months and 1 year. RESULTS: Anatomical regression of extrafoveal polyps was documented on a combination of en face and cross-sectional SSOCTA, 3 months post-laser photocoagulation and anti-vascular endothelial growth factor. Regression of polyps was maintained at the 12-month follow-up visit in all cases. Changes in branching vascular network morphology post-treatment were well-delineated on en face SSOCTA. Swept source optical coherence tomography angiography findings correlated well with the gold standard indocyanine green angiography. CONCLUSION: Swept source optical coherence tomography angiography is an effective noninvasive imaging modality to diagnose and longitudinally follow-up extrafoveal polyps postintervention. Laser photocoagulation with anti-vascular endothelial growth factor achieved regression of polyps in all cases and this was maintained over 12 months.

Computational exploration of the dual role of the phytochemical fortunellin: Antiviral activities against SARS-CoV-2 and immunomodulatory abilities against the host.

Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) infections generate approximately one million virions per day, and the majority of available antivirals are ineffective against it due to the virus's inherent genetic mutability. This necessitates the investigation of concurrent inhibition of multiple SARS-CoV-2 targets. We show that fortunellin (acacetin 7-O-neohesperidoside), a phytochemical, is a promising candidate for preventing and treating coronavirus disease (COVID-19) by targeting multiple key viral target proteins. Fortunellin supports protective immunity while inhibiting pro-inflammatory cytokines and apoptosis pathways and protecting against tissue damage. Fortunellin is a phytochemical found in Gojihwadi kwath, an Indian traditional Ayurvedic formulation with an antiviral activity that is effective in COVID-19 patients. The mechanistic action of its antiviral activity, however, is unknown. The current study comprehensively evaluates the potential therapeutic mechanisms of fortunellin in preventing and treating COVID-19. We have used molecular docking, molecular dynamics simulations, free-energy calculations, host target mining of fortunellin, gene ontology enrichment, pathway analyses, and protein-protein interaction analysis. We discovered that fortunellin reliably binds to key targets that are necessary for viral replication, growth, invasion, and infectivity including Nucleocapsid (N-CTD) (-54.62 kcal/mol), Replicase-monomer at NSP-8 binding site (-34.48 kcal/mol), Replicase-dimer interface (-31.29 kcal/mol), Helicase (-30.02 kcal/mol), Papain-like-protease (-28.12 kcal/mol), 2'-O-methyltransferase (-23.17 kcal/mol), Main-protease (-21.63 kcal/mol), Replicase-monomer at dimer interface (-22.04 kcal/mol), RNA-dependent-RNA-polymerase (-19.98 kcal/mol), Nucleocapsid-NTD (-16.92 kcal/mol), and Endoribonuclease (-16.81 kcal/mol). Furthermore, we identify and evaluate the potential human targets of fortunellin and its effect on the SARS-CoV-2 infected tissues, including normal-human-bronchial-epithelium (NHBE) and lung cells and organoids such as pancreatic, colon, liver, and cornea using a network pharmacology approach. Thus, our findings indicate that fortunellin has a dual role; multi-target antiviral activities against SARS-CoV-2 and immunomodulatory capabilities against the host.

Endogenous fungal endophthalmitis following intensive corticosteroid therapy in severe COVID-19 disease.

Purpose: To report endogenous fungal endophthalmitis, postrecovery from severe COVID-19 infection in otherwise immunocompetent individuals, treated with prolonged systemic steroids. Methods: Retrospective chart review of cases with confirmed and presumed fungal endogenous endophthalmitis, following severe COVID-19 disease, treated at two tertiary care referral eye institutes in North India. Results: Seven eyes of five cases of endogenous fungal endophthalmitis were studied. All cases had been hospitalized for severe COVID-19 pneumonia and had received systemic steroid therapy for an average duration of 42 ± 25.1 days (range 18-80 days). All the cases initially complained of floaters with blurred vision after an average of 6 days (range 1-14 days) following discharge from hospital. They had all been misdiagnosed as noninfectious uveitis by their primary ophthalmologists. All eyes underwent pars plana vitrectomy (PPV) with intravitreal antifungal therapy. Five of the seven eyes grew fungus as the causative organism (Candida sp. in four eyes, Aspergillus sp. in one eye). Postoperatively, all eyes showed control of the infection with a marked reduction in vitreous exudates and improvement in vision. Conclusion: Floaters and blurred vision developed in patients after they recovered from severe COVID-19 infection. They had received prolonged corticosteroid treatment for COVID-19 as well as for suspected noninfectious uveitis. We diagnosed and treated them for endogenous fungal endophthalmitis. All eyes showed anatomical and functional improvement after PPV with antifungal therapy. It is important for ophthalmologists and physicians to be aware of this as prompt treatment could control the infection and salvage vision.

Recent technological advancements in stem cell research for targeted therapeutics.

Stem cells have characteristic features of self-renewal, pluripotency and differentiation, which are responsible for replenishment of tissue or organ. Stem cells are potentiated as therapeutic tool in drug targeting and regenerative medicine-from curing various neurological diseases and malignancies to congenital diseases. These technological advancements have established stem cells as future of medicine. However, due to ethico-social limitations, the use of embryonic stem cells (ESCs) has been avoided, while physiological availability of adult stem cells (ASCs) and induced pluripotent stem cells (iPSCs) has gained appropriate preference. These iPSCs are very much similar to ESCs in terms of their self-renewal and pluripotency. Here, we have summarized the technologies that have established stem cells isolation, their molecular marker and factors responsible for their maintenance. Different cellular (transcription factors, regulatory proteins, miRNA like miRNA-296, miRNA-145, etc.) and extracellular components transcend stem cell fate. Their identification and characterization involve development and efficient utilization of tools like magnetic activated cell sorting (MACS) and fluorescence activated cell sorting (FACS). Some of the technologies have been patented and spin-off's based on them have been commercialized. In conclusion, we present the future scope and possibilities that stem cell technologies behold for us. Graphical abstract Pictorial representation of therapeutic approaches for disease treatment using stem cell technology. Disease-specific adult stem cells are isolated along with niche cells by utilizing tools like FACS/MACS/LCM, etc. Thereafter, cells are reprogrammed through introduction of Yamanaka factors (Oct3/4, Sox2, c-myc, Klf4) to make induced pluripotent stem cell (iPSCs). The disease-specific iPSCs undergo genetic modification after delivery of therapeutic gene through retroviral vehicle. The genetically modified cells are introduced back in person with disease for therapeutic effects. FACS, fluorescence activated cell sorting; MACS, magnetic-activated cell sorting; LCM, laser capture microdissection; Oct3/4, octamer-binding transcription factor 3/4; Sox2, sex determining region Y-box 2; Klf4, Kruppel-like factor 4.

Cytotoxic and apoptotic inducing activity of Amoora rohituka leaf extracts in human breast cancer cells.

BACKGROUND: Amoora rohituka is described in Ayurveda, an Indian traditional system of medicine for management of disorders of blood, diseases of eye, helminthiasis disease, ulcer, liver disorders and splenomegaly. However, the leaves were not reported to have anticancer properties till date. OBJECTIVE: This study was carried out to evaluate the cytotoxic potential of leaf extracts of Amoora rohituka. MATERIALS AND METHODS: The leaves powder was macerated in petroleum ether, ethyl acetate and methanol and evaluated their anticancer activities in vitro. The phytochemical constituents of the active (ethyl acetate) extract were screened by FTIR analysis and phytochemical screening methods. RESULTS: The ethyl acetate extract (RLEA) showed the presence of alkaloids, flavonoids, steroids, tannins, saponins and terpenoids. The RLEA exhibited high cytotoxic effect against human breast cancer cells, MCF-7 (IC50 = 9.81 µg/mL) and induced apoptosis by altering nuclear morphology and DNA laddering. Wound healing assays explained the potency of extract to decrease the cell migration. CONCLUSION: The extract of Amoora rohituka leaves exhibited anticancer activity with less toxicity and it could be used for development of alternative drugs in the treatment of human breast cancer.

Inverted internal limiting membrane (ILM) flap technique for macular hole closure: patient selection and special considerations.

This paper reviews the current status of the newer inverted internal limiting membrane flap technique for macular hole surgery. It gives an overview of the importance of patient selection and special considerations along with variations in technique. It discusses the pathophysiology and how the technique has been an important addition in the armamentarium of vitreoretinal surgeons to attain better anatomical as well as functional results in challenging situations.