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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

Drought mediated physiological and molecular changes in muskmelon (Cucumis melo L.).

Water deficiency up to a certain level and duration leads to a stress condition called drought. It is a multi-dimensional stress causing alteration in the physiological, morphological, biochemical, and molecular traits in plants resulting in improper plant growth and development. Drought is one of the major abiotic stresses responsible for loss of crops including muskmelon (Cucumis melo. L). Muskmelon genotype SC-15, which exhibits high drought resistance as reported in our earlier reports, was exposed to deficient water condition and studied for alteration in physiological, molecular and proteomic profile changes in the leaves. Drought stress results in reduced net photosynthetic rate (Pn), stomatal conductance (Gs) and transpiration (E) rate. With expanded severity of drought, declination recorded in content of total chlorophyll and carotenoid while enhancement observed in phenol content indicating generation of oxidative stress. In contrary, activities of catalase (CAT), superoxide dismutase (SOD), ascorbate peroxidase (APX), and guaiacol (POD) were increased under drought stress. Peptide mass fingerprinting (PMF) showed that drought increased the relative abundance of 38 spots while decreases10 spots of protein. The identified proteins belong to protein synthesis, photosynthesis, nucleotide biosynthesis, stress response, transcription regulation, metabolism, energy and DNA binding. A drought-induced MADS-box transcription factor was identified. The present findings indicate that under drought muskmelon elevates the abundance of defense proteins and suppresses catabolic proteins. The data obtained exhibits possible mechanisms adopted by muskmelon to counter the impacts of drought induced stress.

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.

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.

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.

Hybrid 20/23-G pars plana vitrectomy in endophthalmitis and trauma: a strategic approach.

PURPOSE: To assess the safety and efficacy of hybrid vitreous surgery combining active 23-G ports with a sutured 20-G 6-mm infusion cannula for performing vitrectomy in endophthalmitis and trauma. METHODS: This is a retrospective analysis of 10 eyes with endophthalmitis and 10 eyes with trauma requiring vitreous surgery, having corneal clarity suitable for vitrectomy and best-corrected visual acuity (BCVA) greater than or equal to light perception. All patients underwent hybrid 20/23-G vitrectomy. Intraoperative notes were analyzed for peroperative complications. The BCVA and retinal status at 3 months were analyzed. RESULTS: In the endophthalmitis group, patients ranged from 5 to 85 years of age and showed varied etiologies. Retinal detachment was present in 5 eyes, which received silicone oil tamponade. Mean preoperative logMAR visual acuity (VA) was 2.29 ± 0.45, which improved to 1.10 ± 0.72 at 3 months postoperatively (p<0.001). Trauma cases ranged from 21 to 75 years of age. Retinal detachment was present in 9 out of 10 eyes, all of which received silicone oil tamponade. Preoperative logMAR VA was 2.26 ± 0.71, which improved to 1.33 ± 0.50 postoperatively (p<0.001). At 3 months postsurgery, the retina was attached in all eyes in both groups. CONCLUSIONS: The longer sutured 20-G cannula was easier to visualize through fibrin, exudates, and hemorrhage and did not slip out or enter the suprachoroidal space in any of our cases. The 23-G active ports and vitrectomy cutter enhanced safety and efficacy. Hybrid vitrectomy enabled safe surgery in these difficult cases.

Withania somnifera: Advances and Implementation of Molecular and Tissue Culture Techniques to Enhance Its Application.

Withania somnifera, commonly known as Ashwagandha an important medicinal plant largely used in Ayurvedic and indigenous medicine for over 3,000 years. Being a medicinal plant, dried powder, crude extract as well as purified metabolies of the plant has shown promising therapeutic properties. Withanolides are the principal metabolites, responsible for the medicinal properties of the plant. Availability and amount of particular withanolides differ with tissue type and chemotype and its importance leads to identification characterization of several genes/ enzymes related to withanolide biosynthetic pathway. The modulation in withanolides can be achieved by controlling the environmental conditions like, different tissue culture techniques, altered media compositions, use of elicitors, etc. Among all the in vitro techniques, hairy root culture proved its importance at industrial scale, which also gets benefits due to more accumulation (amount and number) of withanolides in roots tissues of W. somnifera. Use of media compostion and elicitors further enhances the amount of withanolides in hairy roots. Another important modern day technique used for accumulation of desired secondary metabolites is modulating the gene expression by altering environmental conditions (use of different media composition, elicitors, etc.) or through genetic enginnering. Knowing the significance of the gene and the key enzymatic step of the pathway, modulation in withanolide contents can be achieved upto required amount in therapeutic industry. To accomplish maximum productivity through genetic enginnering different means of Withania transformation methods have been developed to obtain maximum transformation efficiency. These standardized transformation procedues have been used to overexpress/silence desired gene in W. somnifera to understand the outcome and succeed with enhanced metabolic production for the ultimate benefit of human race.

Deciphering the Role of microRNAs in BRD4-NUT Fusion Gene Induced NUT Midline Carcinoma.

NUT midline carcinoma (NMC) is a very aggressive and lethal type of squamous epithelial cell cancer caused due to fusion of BRD4 and NUT genes. The gene fusion results into a new fusion protein that promotes oncogenesis. The detailed molecular mechanisms underlying the NMC are still not clear and new findings are urgently required to complement the current efforts. Abnormal microRNAs (miRNA) expression promotes tumour formation by modulating the functional expression of critical genes other than the parent genes involved in tumour cell proliferation or survival. Here, using Insilco methods, miRNA targeting the transcripts of parent genes (BRD4 and NUT) and the BRD4-NUT fusion gene were predicted. We investigated a situation, wherein abnormal miRNA expression in malignant cells could arise due to deletion and fusion of genomic regions encompassing the target site of miRNA genes. A set of 48 dysregulated miRNAs targeting the critical genes other than the parent genes (BRD4 and NUT) was identified. Functional enrichment analysis of KEGG pathways of target genes of these Ex-miRNAs implicates their role in cancer pathways. Amplification in the expression level of these miRNAs can be used for NMC diagnosis and prognosis.