Microbial influencers and cotton leaf curl disease (CLCuD) susceptibility: a network perspective.

Biotic stresses, such as plant viruses, e.g., cotton leaf curl virus (CLCuV), can alter root-associated and leaf-associated microbial diversities in plants. There are complex ecological dynamics at play, with each microbe contributing to a multitude of biotic and abiotic interactions, thus deciding the stability of the plant's ecosystem in response to the disease. Deciphering these networks of interactions is a challenging task. The inferential research in microbiome is also at a nascent stage, often constrained by the underlying analytical assumptions and the limitations with respect to the depth of sequencing. There is also no real consensus on network-wide statistics to identify the influential microbial players in a network. Guided by the latest developments in network science, including recently published metrics such as Integrated View of Influence (IVI) and some other centrality measures, this study provides an exposé of the most influential nodes in the rhizospheric and phyllospheric microbial networks of the cotton leaf curl disease (CLCuD) susceptible, partially tolerant, and resistant cotton varieties. It is evident from our results that the CLCuD-resistant Gossypium arboreum possesses an equal share of keystone species, which helps it to withstand ecological pressures. In the resistant variety, the phyllosphere harbors the most influential nodes, whereas in the susceptible variety, they are present in the rhizosphere. Based on hubness score, spreading score, and IVI, the top 10 occurring keystone species in the FDH-228 (resistant) variety include Actinokineospora, Cohnella, Thermobacillus, Clostridium, Desulfofarcimen, and MDD-D21. Elusimicrobia, Clostridium-sensu-stricto_12, Candidatus woesebacteria, and Dyella were identified as the most influential nodes in the PFV-1 (partially tolerant) variety. In the PFV-2 (susceptible) variety, the keystone species were identified as Georginia, Nesterenkonia, Elusimicrobia MVP-88, Acetivibrio, Tepedisphaerales, Chelatococcus, Nitrosospira, and RCP2-54. This concept deciphers the diseased and healthy plant's response to viral disease, which may be microbially mediated.

ULTRASOUND BIOMICROSCOPY AND ECHOGENIC EXTERNAL MARKER ASSISTED INTRAOCULAR FOREIGN BODY REMOVAL.

PURPOSE: We report a novel method of intraoperative localization of a retained anterior intraocular foreign body (IOFB), using a combination of ultrasound biomicrosopy and an insulated needle. METHODS: A retrospective case report of a 56-year-old man who presented with a right eye IOFB. RESULTS: On presentation, vision was decreased in the right eye to count fingers with a small subconjunctival hemorrhage, but no other signs of a penetrating laceration. Orbital CT revealed an IOFB, and the initial vitrectomy failed to retrieve the IOFB. Then, during the subsequent vitrectomy, using an ultrasound compatible needle and an ultrasound biomicrosopy, we were able to precisely locate and remove the small anterior IOFB. CONCLUSION: Retained IOFBs can lead to severe irreversible vision loss if not promptly removed. Ancillary imaging modalities and localization techniques can help locate occult IOFBs in difficult cases.

Cotton microbiome profiling and Cotton Leaf Curl Disease (CLCuD) suppression through microbial consortia associated with Gossypium arboreum.

The failure of breeding strategies has caused scientists to shift to other means where the new approach involves exploring the microbiome to modulate plant defense mechanisms against Cotton Leaf Curl Disease (CLCuD). The cotton microbiome of CLCuD-resistant varieties may harbor a multitude of bacterial genera that significantly contribute to disease resistance and provide information on metabolic pathways that differ between the susceptible and resistant varieties. The current study explores the microbiome of CLCuD-susceptible Gossypium hirsutum and CLCuD-resistant Gossypium arboreum using 16 S rRNA gene amplification for the leaf endophyte, leaf epiphyte, rhizosphere, and root endophyte of the two cotton species. This revealed that Pseudomonas inhabited the rhizosphere while Bacillus was predominantly found in the phyllosphere of CLCuV-resistant G. arboreum. Using salicylic acid-producing Serratia spp. and Fictibacillus spp. isolated from CLCuD-resistant G. arboreum, and guided by our analyses, we have successfully suppressed CLCuD in the susceptible G. hirsutum through pot assays. The applied strains exhibited less than 10% CLCuD incidence as compared to control group where it was 40% at 40 days post viral inoculation. Through detailed analytics, we have successfully demonstrated that the applied microbes serve as a biocontrol agent to suppress viral disease in Cotton.

Bilateral Cystoid Macular Edema Following Bimatoprost Implants.

PURPOSE: To report the development of bilateral cystoid macular edema (CME) following bimatoprost implant (Durysta) injections in both eyes to treat primary open angle glaucoma (POAG). METHODS: Case Report. RESULTS: A 93-year-old woman with a history of POAG received bimatoprost implant (Durysta) injections in both eyes four weeks apart. The patient subsequently developed progressively decreased visual acuity in both eyes due to bilateral CME, which improved with topical corticosteroid therapy. CONCLUSIONS: Bimatoprost implant (Durysta) can cause CME in susceptible individuals. Patients who received the implant should be assessed for the presence of CME following any decline in visual acuity, particularly in high-risk patients.

Studies on biochemical, oxidative and genotoxicity alterations following vas blockage with reversible inhibition of sperm under guidance and reversal in rats.

CONTEXT: Vas obstruction with reversible inhibition of sperm under guidance (RISUG) for contraception and its reversal, may cause oxidative stress or inimical effects on male reproductive functions. OBJECTIVE: To evaluate the biochemical and genotoxicity at the level of reactive oxygen species (ROS) following vas occlusion with RISUG and its reversal by Dimethyl sulphoxide (DMSO) and 5% NaHCO3 in Wistar albino rats. SETTINGS AND DESIGN: Animals were divided into seven groups (n = 10), namely sham-operated control, short-term vas occlusion with RISUG for 90 days, reversal with DMSO and 5% NaHCO3, long-term vas occlusion with RISUG for 360 days, reversal with DMSO and 5% NaHCO3. MATERIALS AND METHODS: Biochemical markers in reproductive tissues, hematology, serum biochemistry, serum electrolytes and ROS measuring indicators, e.g., lipid peroxidation, superoxide dismutase, catalase, glutathione peroxidase, and glutathione S-transferase were examined. STATISTICAL ANALYSIS USED: One-way analysis of variance test was performed for analyses of data obtained in this study using the SPSS 10.0 software (SPSS Inc., Chicago, IL, USA). RESULTS: The tissue and clinical chemistry did not show appreciable alterations in RISUG injected and reversal Groups (II-VII) as compared to sham control. The genotoxicity and various ROS markers fluctuated within control limits following short- and long-term vas occlusion and reversal. CONCLUSIONS: The study suggested that the reversal procedures, following RISUG contraception, were not associated with any kind of toxicological manifestations.

Reversible inhibition of sperm under guidance as an intratubular and reversible contraception in female rats: An experimental study.

BACKGROUND: Reversible inhibition of sperm under guidance ("RISUGⓇ") is a promising intravasal male contraceptive. OBJECTIVE: An exploratory study was conducted with a concept of non-invasive, transcervical, single-intervention and reversible contraception using RISUGⓇ in females. MATERIALS AND METHODS: In this experimental study, 60 adult Wistar albino female rats weighing 150-155 g, 3-4 months old were divided into four groups: group I: sham-operated control; group II: tubal occlusion with RISUG for 90 days; group III: tubal occlusion with RISUGⓇ for 90 days and reversal with dimethyl sulphoxide and group IV: tubal occlusion with RISUGⓇ for 90 days and reversal with 5% NaHCO. Animals were subjected to bilateral fallopian tube occlusion with RISUGⓇ and reversal with DMSO and NaHCO 3 . The estrous cycle, fertility and histology of fallopian tube were evaluated. RESULTS: Group I showed 100% fertility during all mating schedules. Animals of experimental groups indicated positive mating, but 0% fertility was evident following 30, 60, and 90 days of tubal occlusion. However, after reversal, fertility steadily increased to normalcy in groups III (50% at 45 days, 80% at 105 days, 100% at 150 and 195 days) and IV (70% at 45 and 105 days, 100% at 150 and 195 days) animals. Group II illustrated disorganized inner cell linings and eosinated RISUGⓇimplant-filled lumen. Reversal groups (III and IV) revealed complete restoration of cellular histo-architecture. Regular estrous cycle was noticed in all experimental groups. CONCLUSION: RISUGⓇ is suitable for single intervention, intratubular, reversible contraception in female rats.

RISUG® as a male contraceptive: journey from bench to bedside.

Even after decades of research men still lack reliable and reversible contraceptive methods comparable to female methods of contraception. Traditional methods of male contraception present a high failure rate and also involve high risk both when used for contraception and for protection against sexually transmitted diseases. Various chemical, hormonal, immunological, vas based and herbal methods of contraception have been examined by scientists world over during the past four decades. Among the possible lead approaches, exogenous hormonal contraception, either alone or in combination with progesterone or antiandrogen, is being viewed at low profile because of their insufficiency in inducing uniform suppression of spermatogenesis and steroid related long term complications. As an alternative to vasectomy, among various intravasal devices being examined, RISUG® (Reversible Inhibition of Sperm Under Guidance), a co-polymer of styrene and maleic anhydride offers long term contraception with safety, efficacy and it can be delivered by no-scalpel injection. Thus it is the only male contraceptive procedure currently under Phase- III Clinical Trial. The non-invasive reversal technique, successfully demonstrated in langur monkeys and functional reversal achieved with dimethyl sulphoxide (DMSO) and sodium bicarbonate (NaHCO3) in rats and rabbits with safety at F1 generation (first filial generation) have projected RISUG® as a better alternative to vasectomy. In this narrative review we revisit the long journey of RISUG® beginning with formulation on a bench towards reaching the market as a safe and effective contraceptive method, discussing various milestones and roadblocks of this expedition awaiting the mandatory regulatory clearance from the Government of India. Successful completion of ongoing phase III clinical trials with demonstration of reversal in human volunteers will give an indigenously developed male contraceptive to the world.

Malgré plusieurs décennies de recherche, il manque toujours pour les hommes des méthodes de contraception fiables et réversibles qui soient comparables aux méthodes de contraception féminine. Les méthodes de contraception masculine traditionnelles ont un taux d'échec élevé ; elles sont aussi à risque lors d'utilisations à la fois comme contraceptif et comme protection contre les infections sexuellement transmissibles. Au cours des 40 dernières années, le monde scientifique a évalué différentes méthodes de contraception masculine basées sur des approches chimiques, hormonales, immunologiques, déférentielles ou à base de plante.Parmi les pistes possibles d'approche, la contraception par apport d'hormone exogène, soit seule soit associée à un progestatif ou à un anti androgène, est actuellement perçue comme ayant un faible profil en raison de son incapacité à induire une suppression uniforme de la spermatogenèse et des complications à long termes des stéroïdes.Parmi les alternatives à la vasectomie, plusieurs dispositifs intra déférentiels ont été évalués dont le RISUG® (Reversible Inhibition of Sperm Under Guidance). Ce dernier est un copolymère de styrène et d'anhydride maléique qui offre une contraception de longue durée avec innocuité et efficacité, pouvant être mise en place par injection sans scalpel. C'est ainsi actuellement la seule procédure contraceptive masculine pour laquelle un essai clinique de Phase-III est en cours. La technique de réversibilité sans scalpel, démontrée avec succès chez le singe Langur, ainsi que la réversibilité fonctionnelle par le sulfoxyde de diméthyle (DMSO) et le bicarbonate de sodium (NaHCO3) confirmée chez le rat et le lapin, sans atteintes sur la génération F1 (première génération de petits), ont fait apparaître le RISUG® comme une meilleure alternative à la vasectomie.Dans cette revue narrative, nous réévaluons le long chemin du RISUG® depuis une formulation de paillasse de laboratoire jusqu'à sa mise sur le marché comme méthode de contraception sans risque et efficace, en discutant les différents jalons et obstacles rencontrés au cours de cette expédition dans l'attente de l'autorisation réglementaire obligatoire du Gouvernement Indien. Le succès des essais de Phase-III en cours par la preuve d'une réversibilité chez des hommes volontaires apportera au Monde une méthode de contraception développée localement.

Toxicity and Mutagenicity Evaluation Following RISUG Contraception Reversal in Rats.

Reestablishment of fertility, after a male contraceptive method, is of great concern. In this context, RISUG (Reversible Inhibition of Sperm Under Guidance) has been evaluated for its mutagenicity following reversibility with dimethyl sulfoxide (DMSO)/sodium bicarbonate (NaHCO3) in Wistar albino rats. Animals were divided into 7 groups, namely, sham-operated control, vas occlusion with RISUG for 90 and 360 days, reversal with DMSO and NaHCO3 after 90 and 360 days, respectively. The testis, cauda epididymis, cauda epididymal spermatozoa, and serum were evaluated for apoptosis and hormonal status through various assays. RISUG was subjected to Ames test at dose levels of 10, 50, and 100 µL. Results of terminal deoxynucleotidyl transferase nick end labeling and caspase-3 assays in testes and cauda epididymis revealed that the percentage of positive cells in the experimental groups was comparable to sham-operated control. Annexin V assay in cauda epididymal spermatozoa showed slight elevation in group II ( P < 0.05), whereas in the remaining groups, minimum numbers of positive sperms were found. Hormone profile, namely, testosterone, prolactin, cortisol, prostate-specific antigen, and sperm antibody concentration, remained unchanged. In Ames test, no significant increase was observed in the number of revertant colonies on plates containing RISUG in the presence and absence of S9 mix in all 3 strains. Therefore, the reversal of RISUG-induced contraception by solvent vehicle DMSO/NaHCO3 was successful without any toxicity at the cellular levels.

Contraception with RISUG® and functional reversal through DMSO and NaHCO3 in male rabbits.

The study aimed to evaluate reversal of short- and long-term vas occlusion with reversible inhibition of sperm under guidance (RISUG) using dimethyl sulfoxide (DMSO) and sodium bicarbonate (NaHCO3) in male rabbits (Oryctolagus cuniculus). Animals were divided into seven groups containing five animals each. Fortnightly, semen analysis revealed that sperm concentration and output steadily declined after vas occlusion and complete azoospermia was attained at 30-60 days postinjection. Spermatozoa reappeared at 60-75 days of reversal and normozoospermia was noticed between 135 days and 150 days in the reversal groups. All spermatozoa were found nonmotile prior to azoospermia and a gradual recovery in sperm motility was observed between 105 days and 135 days of reversal. A significant decline in viability of sperms was noticed during vas occlusion up to 30-60 days which recovered at 60-75 days postreversal and normalized by 75-105 days in the reversal groups. A significant enhancement in the sperm abnormalities was recorded in all vas occluded animals as well as those in initial periods of reversal. Other parameters, namely, semen volume, ejaculation time, pH, color, and consistency, remained unaltered during all phases of the study. Fertility test, at the intervals of 15 days, demonstrated that animals exhibited complete sterility during the entire period of vas occlusion. A gradual recovery in fertility was observed with the appearance of spermatozoa following vas occlusion reversal and 100% fertility was observed following 135-150 days of reversal. F1 progeny of reversed animals was found normal. The results suggest that reversal with DMSO or NaHCO3 is feasible, with normal progeny, following short- and long-term contraception.