Comparative proteome analysis of Spodoptera litura-infested Zea mays reveals a robust defense strategy targeting insect peritrophic membrane.

Herbivorous insects such as Spodoptera litura, pose a constant threat to agricultural crops. The incompetence of contemporary pest management tools and techniques stipulates unravelling of molecular dogma, that drives pest-plant interaction. From our previous observations, we inferred that despite being a voracious polyphagous herbivore, S. litura growth and adaptability is severely hampered on maize foliage diet. In this investigation we explored further and demonstrated the impact of maize diet on the insect gut peritrophic membrane (PM, a crucial membrane involved in compartmentalizing digestive events and absorption of nutrients), its structural analysis using scanning electron microscopy (SEM) revealed damaged and perforated PM. Further, this study delves into the intricate resistance mechanism adapted by Z. mays against S. litura by conducting a comparative proteome analysis. We have detected 345 differentially abundant proteins (DAPs) at p < 0.05 and fold change ≥1. The DAPs were categorized as plant defense, secondary metabolite synthesis, redox homeostasis, cytoskeleton/cell wall biosynthesis, primary metabolism, transport and molecular processes. We remarkably report differential expression of proteolysis- and defense-related proteins that have potential to target insect gut, digestion and absorption of nutrients. Our findings contribute to a deeper understanding of the molecular dynamics governing maize resistance against S. litura. Understanding of such intricate molecular dialogues at these interfaces could provide valuable information on the arms race between plants and herbivores, it may pave the way for innovative pest management strategies.

Discovery and multimerization of cross-reactive single-domain antibodies against SARS-like viruses to enhance potency and address emerging SARS-CoV-2 variants.

Coronaviruses have been the causative agent of three epidemics and pandemics in the past two decades, including the ongoing COVID-19 pandemic. A broadly-neutralizing coronavirus therapeutic is desirable not only to prevent and treat COVID-19, but also to provide protection for high-risk populations against future emergent coronaviruses. As all coronaviruses use spike proteins on the viral surface to enter the host cells, and these spike proteins share sequence and structural homology, we set out to discover cross-reactive biologic agents targeting the spike protein to block viral entry. Through llama immunization campaigns, we have identified single domain antibodies (VHHs) that are cross-reactive against multiple emergent coronaviruses (SARS-CoV, SARS-CoV-2, and MERS). Importantly, a number of these antibodies show sub-nanomolar potency towards all SARS-like viruses including emergent CoV-2 variants. We identified nine distinct epitopes on the spike protein targeted by these VHHs. Further, by engineering VHHs targeting distinct, conserved epitopes into multi-valent formats, we significantly enhanced their neutralization potencies compared to the corresponding VHH cocktails. We believe this approach is ideally suited to address both emerging SARS-CoV-2 variants during the current pandemic as well as potential future pandemics caused by SARS-like coronaviruses.

Hexamerization of Anti-SARS CoV IgG1 Antibodies Improves Neutralization Capacity.

The SARS-CoV-2 pandemic and particularly the emerging variants have deepened the need for widely available therapeutic options. We have demonstrated that hexamer-enhancing mutations in the Fc region of anti-SARS-CoV IgG antibodies lead to a noticeable improvement in IC50 in both pseudo and live virus neutralization assay compared to parental molecules. We also show that hexamer-enhancing mutants improve C1q binding to target surface. To our knowledge, this is the first time this format has been explored for application in viral neutralization and the studies provide proof-of-concept for the use of hexamer-enhanced IgG1 molecules as potential anti-viral therapeutics.

Molecular Rationale of Insect-Microbes Symbiosis-From Insect Behaviour to Mechanism.

Insects nurture a panoply of microbial populations that are often obligatory and exist mutually with their hosts. Symbionts not only impact their host fitness but also shape the trajectory of their phenotype. This co-constructed niche successfully evolved long in the past to mark advanced ecological specialization. The resident microbes regulate insect nutrition by controlling their host plant specialization and immunity. It enhances the host fitness and performance by detoxifying toxins secreted by the predators and abstains them. The profound effect of a microbial population on insect physiology and behaviour is exploited to understand the host-microbial system in diverse taxa. Emergent research of insect-associated microbes has revealed their potential to modulate insect brain functions and, ultimately, control their behaviours, including social interactions. The revelation of the gut microbiota-brain axis has now unravelled insects as a cost-effective potential model to study neurodegenerative disorders and behavioural dysfunctions in humans. This article reviewed our knowledge about the insect-microbial system, an exquisite network of interactions operating between insects and microbes, its mechanistic insight that holds intricate multi-organismal systems in harmony, and its future perspectives. The demystification of molecular networks governing insect-microbial symbiosis will reveal the perplexing behaviours of insects that could be utilized in managing insect pests.

Dynamics of Zea mays transcriptome in response to a polyphagous herbivore, Spodoptera litura.

Zea mays defense response is well-crafted according to the physical and chemical weapons utilized by their invaders during the coevolutionary period. Maize plants employ diversified defense strategies and alter the spatiotemporal distribution of several classes of defensive compounds to affect insect herbivore performance. However, only little knowledge is available about the defense orchestration of maize in response to Spodoptera litura, a voracious Noctuidae pest. In order to decipher the defense status of Zea mays (African tall variety) against S. litura, a comparative feeding bioassay was executed, which revealed reduced performance of the herbivore on maize. In order to understand the molecular mechanism behind maize tolerance against S. litura, a microarray-based genome-wide expression analysis was performed. The comparative analysis displayed 792 differentially expressed genes (DEGs), wherein 357 genes were upregulated and 435 genes were downregulated at fold change ≥ 2 and p value ≤ 0.05. The upregulated genes were identified and categorized as defense-related, oxidative stress-related, transcription regulatory genes, protein synthesis genes, phytohormone-related, and primary and secondary metabolism-related. In contrast, downregulated genes were mainly associated with plant growth and development, indicating a balance of growth and defense response and utilization of a highly evolved C-diversion response were noticed. Maize plants showed better tolerance against herbivory and maintained its fitness using a combinatorial strategy. This peculiar response of Zea mays against S. litura offers an excellent possibility of managing polyphagous pests by spicing up the plant's defensive response with tolerance mechanism.

Myeloid cell leukemia 1 (MCL-1): Structural characteristics and application in cancer therapy.

Apoptosis, a major hallmark of cancer cells, regulates cellular fate and homeostasis. BCL-2 (B-cell CLL/Lymphoma 2) protein family is popularly known to mediate the intrinsic mode of apoptosis, of which MCL-1 is a crucial member. Myeloid cell leukemia 1 (MCL-1) is an anti-apoptotic oncoprotein and one of the most investigated members of the BCL-2 family. It is commonly known to be genetically altered, aberrantly overexpressed, and primarily associated with drug resistance in various human cancers. Recent advancements in the development of selective MCL-1 inhibitors and evaluating their effectiveness in cancer treatment establish its popularity as a molecular target. The overall aim is the selective induction of apoptosis in cancer cells by using a single or combination of BCL-2 family inhibitors. Delineating the precise molecular mechanisms associated with MCL-1-mediated cancer progression will certainly improve the efficacy of clinical interventions aimed at MCL-1 and hence patient survival. This review is structured to highlight the structural characteristics of MCL-1, its specific interactions with NOXA, MCL-1-regulatory microRNAs, and at the same time focus on the emerging therapeutic strategies targeting our protein of interest (MCL-1), alone or in combination with other treatments.

Associations between parents' exposure to a multisectoral programme and infant and young child feeding practices in Nepal.

In Nepal, an at-scale, multisectoral programme-Suaahara (2011-2023)-aims to improve nutrition behaviours. Suaahara II (2016-2023) transitioned from a mother/child dyad focus to explicitly targeting all family members. Evidence is scant, however, regarding how exposure by men to social and behaviour change interventions relates to nutrition outcomes. This study uses a 2019 cross-sectional monitoring dataset to test associations between maternal and male household head exposure to Suaahara II interventions (interacting with a frontline worker, participating in a community event or listening to the Bhanchhin Aama radio programme) and adoption of three infant and young child feeding practices: minimum dietary diversity, minimum acceptable diet and sick child feeding, in households with a child under 2 years (n = 1827). Maternal exposure to Suaahara II had a positive association with minimum dietary diversity (OR: 1.71, 95% CI [1.27, 2.28], P < 0.001), minimum acceptable diet (OR: 1.60, 95% CI [1.19, 2.14], P = 0.002) and increased feeding to a sick child (OR: 2.11, 95% CI [1.41, 3.17], P < 0.001). Male household head exposure was only associated with increased feeding to a sick child (OR: 2.21, 95% CI [1.27, 3.84], P = 0.005). Among households with an exposed mother, having an exposed male household head nearly tripled the odds of appropriate sick child feeding (OR: 2.90, 95% CI [1.57, 5.34], P = 0.001) but was not significantly associated with the other two outcomes. These findings suggest that the relationships between exposure to nutrition programmes and outcomes are complex and further research is needed to understand variation by family member, behavioural outcome and context.

Molecular Modeling of Chemosensory Protein 3 from Spodoptera litura and Its Binding Property with Plant Defensive Metabolites.

Chemosensory perception in insects involves a broad set of chemosensory proteins (CSPs) that identify the bouquet of chemical compounds present in the external environment and regulate specific behaviors. The current study is focused on the Spodoptera litura (Fabricius) chemosensory-related protein, SlitCSP3, a midgut-expressed CSP, which demonstrates differential gene expression upon different diet intake. There is an intriguing possibility that SlitCSP3 can perceive food-derived chemical signals and modulate insect feeding behavior. We predicted the three-dimensional structure of SlitCSP3 and subsequently performed an accelerated molecular dynamics (aMD) simulation of the best-modeled structure. SlitCSP3 structure has six α-helices arranged as a prism and a hydrophobic binding pocket predominated by leucine and isoleucine. We analyzed the interaction of selected host plant metabolites with the modeled structure of SlitCSP3. Out of two predicted binding pockets in SlitCSP3, the plant-derived defensive metabolites 2-b-D-glucopyranosyloxy-4-hydroxy-7-methoxy-1, 4-benzoxazin-3-one (DIMBOA), 6-Methoxy-2-benzoxazolinone (MBOA), and nicotine were found to interact preferably to the hydrophobic site 1, compared to site 2. The current study provides the potential role of CSPs in recognizing food-derived chemical signals, host-plant specialization, and adaptation to the varied ecosystem. Our work opens new perspectives in designing novel pest-management strategies. It can be further used in the development of CSP-based advanced biosensors.

Protease inhibitors: recent advancement in its usage as a potential biocontrol agent for insect pest management.

Plant-derived protease inhibitors (PIs) are a promising defensin for crop improvement and insect pest management. Although agronomist made significant efforts in utilizing PIs for managing insect pests, the potentials of PIs are still obscured. Insect ability to compensate nutrient starvation induced by dietary PI feeding using different strategies, that is, overexpression of PI-sensitive protease, expression of PI-insensitive proteases, degradation of PI, has made this innumerable collection of PIs worthless. A practical challenge for agronomist is to identify potent PI candidates, to limit insect compensatory responses and to elucidate insect compensatory and resistance mechanisms activated upon herbivory. This knowledge could be further efficiently utilized to identify potential targets for RNAi-mediated pest control. These vital genes of insects could be functionally annotated using the advanced gene-editing technique, CRISPR/Cas9. Contemporary research is exploiting different in silico and modern molecular biology techniques to utilize PIs in controlling insect pests efficiently. This review is structured to update recent advancements in this field, along with its chronological background.

Effect of 2 Novel Sustained-release Drug Release Systems on Bleb Fibrosis: An In Vivo Trabeculectomy Study in a Rabbit Model.

PRECIS: Rabbit model studies suggested better morphology blebs with equal intraocular pressure (IOP) efficacy as a standard mitomycin C (MMC) trabeculectomy using a novel slow-release drug delivery antifibrotic system delivering small quantities of MMC and 5-fluorouracil (5-FU). PURPOSE: To evaluate 2 different concentrations of biodegradable poly(lactic-co-glycolic acid) (PLGA) system with 5-FU and MMC (ElutiGLASS) for their ability to reduce fibrosis and compare the results with standard trabeculectomy with MMC in a rabbit model. MATERIALS AND METHODS: New Zealand albino rabbits (19) were divided into 3 groups (A, B, C) and standard trabeculectomy operation was performed in the right eye of each rabbit.Group (A) had trabeculectomy with MMC (0.4 mg/mL) applied using a Weck cell sponge; (B) trabeculectomy with slow-release ElutiGLASS (0.23 mg, 5-FU/0.33 µg MMC released over 23 to 30 d); (C) trabeculectomy with rapid release ElutiGLASS (0.45 mg of 5-FU/0.65 µg MMC, released over 5 to 7 d). The rabbits were followed for 3 months before euthanasia. RESULTS: Bleb morphology, vascularity, and fibrosis were less pronounced in groups B and C when compared with group A at 3 months. Group B appears to have a lower and more diffuse bleb appearance compared with the other 2 groups with honeycomb appearance on both clinical examination and ultrasound biomicroscopy imaging with higher percentage of maintained bleb space (83%), less fibrosis than group A while maintaining the same low inflammation score as the other 2 groups on histology. At 3 months, the PLGA polymer had completely disappeared in all rabbits. There were no statistical differences in the degree of IOP reduction or histologic inflammation, among the 3 groups. CONCLUSIONS: We successfully created a sustained-release antifibrotic drug delivery system that delivered known dosage of the drugs at doses that are significantly lower than the current standard, and resulted in less fibrosis while maintaining a healthy bleb and equal reduction of IOP. TRANSLATIONAL RELEVANCE: These results are supportive of the antifibrotic effect of the slow-release drug delivery system used in conjunction with trabeculectomy, thus paving the way for human pilot studies to improve and simplify existing surgical techniques for filtering surgeries in glaucoma.

In silico prediction of active site and in vitro DNase and RNase activities of Helicoverpa-inducible pathogenesis related-4 protein from Cicer arietinum.

Plants are endowed with an innate immune system, which enables them to protect themselves from pest and pathogen. The participation of pathogenesis-related (PR) proteins is one of the most crucial events of inducible plant defense response. Herein, we report the characterization of CaHaPR-4, a Helicoverpa-inducible class II PR-4 protein from chickpea. Bioinformatic analysis of CaHaPR-4 protein indicated the presence of a signal peptide, barwin domain but it lacks the chitin-binding site/hevein domain. The recombinant CaHaPR-4 is bestowed with RNase and bivalent ion-dependent DNase activity. Further, the RNA and DNA binding sites were identified and confirmed by analyzing interactions between mutated CaHaPR-4 with the altered active site and ribonuclease inhibitor, 5'ADP and DNase inhibitor, 2­nitro­5­thiocyanobenzoic acid (NTCB) using 3D modeling and docking studies. Moreover, CaHaPR-4 shows antifungal activity as well as growth inhibiting properties against neonatal podborer larvae. To the best of our knowledge, this is the first report of a PR-4 showing RNase, DNase, antifungal and most importantly insect growth inhibiting properties against Helicoverpa armigera simultaneously.

A case of hypercalcaemia in an immunocompetent patient with Mycobacterium avium intracellulare.

Mycobacterium avium intracellulare infection or colonization should be considered in the differential diagnosis of hypercalcemia, especially in immunocompromised individuals, in the appropriate clinical context.

Quadruplet pregnancy following spontaneous conception: a rare case report.

Quadruplets are a set of four offspring born at one birth which can be fraternal (multizygotic), identical (monozygotic) or a combination of both. Multizygotic quadruplets occur from fertilisation of four different sets of ovum and sperm. Monozygotic multiples are the result of a fertilized egg that splits into two or more embryos. Multizygotic quadruplets can be all male, all female, or a combination of both while monozygotic quadruplets will always be of the same gender. Here we present a case of 32-year-old G4P3L0 with previous history of three term intrauterine foetal death (IUFD) at 27 wk of gestation. After evaluation, she was found to carry three live foetuses with an IUFD. She was managed conservatively till 35 wk of gestation with regular monitoring of coagulation parameters. Elective caesarean section was done at 35 wk and three live female babies and one male still born were delivered. This case is unique due to the fact that our patient conceived a multizygotic quadruplet pregnancy spontaneously following a bad obstetric history (3 previous term intrauterine fetal deaths). One fetus died in utero and the pregnancy continued successfully resulting in 3 live born healthy babies.

Genetics of fetal hemoglobin in tribal Indian patients with sickle cell anemia.

India tops the list of countries with sickle cell disease (SCD) with an estimated 44,000 live births in 2010 and a prevalence of 10%-33%. In the present study, the first from India, we have investigated the effect of genetic variants in the BCL11A, the HMIP (HBS1L-MYB intergenic polymorphism) locus, in addition to the HBB locus, which are known to be associated with fetal hemoglobin (HbF) levels, a major modulator of the disease phenotype. The present study was conducted on 240 individuals with SCD and 60 with sickle cell trait. Genotyping was performed for the BCL11A rs11886868 and rs34211119; HMIP rs9399137, rs189600565, rs7776196, rs34778774, and rs53293029; HBG2 Xmn1 polymorphism rs7482144; and -68C > T HBD promoter polymorphism. All the 3 quantitative trait loci were associated with HbF levels in Indian patients with SCD. The highest difference was seen in the Xmn1 single-nucleotide polymorphism, which accounted for 11% of the trait variance, the BCL11A rs11886868 for 3.65%, whereas the HMIP rs9399137 for 3.8%. The present study indicates the BCL11A, HMIP, and ß-globin region to be associated with increased HbF levels in Indian patient. Further interrogation of these genotypes with respect to pain crisis is warranted in this population, which may help in prognostication, as also a genome-wide association study, which may help uncover new loci controlling HbF levels.

Hypoplasia, pseudocoarctation and coarctation of the aorta - a systematic review.

Aortic arch abnormalities are uncommon and may be seen in association with other congenital cardiac anomalies. Coarctation, pseudocoarctation and hypoplastic aortic arch are known aortic arch abnormalities, with the former being well studied, whilst for the latter two, much less is known. There are similarities and differences that are important to distinguish among these three conditions in order to avoid errors in diagnosis that may result in unnecessary investigations, which may in turn result in physical or emotional harm to the patient. For this reason, we present a systematic review of the published literature providing an evidence-based overview that may be helpful to clinicians when faced with this diagnostic dilemma.

In vitro characterization of the anti-PD-1 antibody nivolumab, BMS-936558, and in vivo toxicology in non-human primates.

The programmed death-1 (PD-1) receptor serves as an immunologic checkpoint, limiting bystander tissue damage and preventing the development of autoimmunity during inflammatory responses. PD-1 is expressed by activated T cells and downmodulates T-cell effector functions upon binding to its ligands, PD-L1 and PD-L2, on antigen-presenting cells. In patients with cancer, the expression of PD-1 on tumor-infiltrating lymphocytes and its interaction with the ligands on tumor and immune cells in the tumor microenvironment undermine antitumor immunity and support its rationale for PD-1 blockade in cancer immunotherapy. This report details the development and characterization of nivolumab, a fully human IgG4 (S228P) anti-PD-1 receptor-blocking monoclonal antibody. Nivolumab binds to PD-1 with high affinity and specificity, and effectively inhibits the interaction between PD-1 and its ligands. In vitro assays demonstrated the ability of nivolumab to potently enhance T-cell responses and cytokine production in the mixed lymphocyte reaction and superantigen or cytomegalovirus stimulation assays. No in vitro antibody-dependent cell-mediated or complement-dependent cytotoxicity was observed with the use of nivolumab and activated T cells as targets. Nivolumab treatment did not induce adverse immune-related events when given to cynomolgus macaques at high concentrations, independent of circulating anti-nivolumab antibodies where observed. These data provide a comprehensive preclinical characterization of nivolumab, for which antitumor activity and safety have been demonstrated in human clinical trials in various solid tumors.

Interferon-α and pericardial injury: a case report and literature review.

Interferon- α (IFN-α) alone or in combination with other chemotherapeutic agents has been used in the management of many malignant and non-malignant conditions. Pericarditis with or without pericardial effusion has been reported with IFN-α therapy, and available literature is limited to case reports. Pericardial constriction after interferon use has not been described in the published literature to date. We performed a systematic review of literature to address the demographic features, clinical presentation, diagnosis, treatment and outcome of interferon-related pericardial injury.

Effect of 82% Lactic Acid in Treatment of Melasma.

Melasma is an acquired, chronic, and symmetrical hypermelanosis, characterized by brown patches of variable darkness on sun exposed areas of body. There are numerous modalities of treatment currently in use for this disease, of which the chemical peeling is very commonly used. Therefore, the present work was done to see the effect of 82% lactic acid peel in the treatment of melasma. A total number of 20 patients of either sex attending the OPD of dermatology department with clinically evident melasma were included in the study. 82% Lactic acid peel was applied on the face for 12 weeks in each patient. Patients were evaluated clinically and photographically at various intervals and in follow-up till 24 weeks. Assessment of patient satisfaction and side effects were also noted. All the subjects completed the study. Application of this peel for 12 weeks significantly decreased the melasma area severity index score and also melasma severity scale score. Patient and physician analogue scales also showed the improvement by the treatment. Regarding the adverse effects, burning sensation was the only side effect noted in our study. In conclusion, 82% lactic acid peel is well tolerated and can be used for the treatment of melasma.

Ameliorative Potential of Quercetin Against Paracetamol-induced Oxidative Stress in Mice Blood.

The aim of the present study was to evaluate the ameliorative potential of quercetin (QC) against paracetamol (PCM)-induced oxidative stress and biochemical alterations in mice blood. A total of 36 mice were randomly allocated into six groups, six mice in each. Group I served as healthy controls, while groups II and III were administered with N-acetylcysteine (NAC) and QC alone respectively. Group IV was administered with PCM alone. Groups V and VI were administered with PCM on day 0 followed by NAC and QC, respectively, for 6 consecutive days. On day 7(th) blood samples were obtained and subjected for the assays of oxidative stress and serum biochemical panels. Erythrocytic lipid peroxides contents of alone PCM-intoxicated mice were significantly higher, while reduced glutathione contents were found to be significantly lower in comparison with the healthy controls. The activities of antioxidant enzymes were also found to be singnificantly lower in these mice. Additionally, significantly increased activities of serum aspartate transaminase, alanine transaminase and alkaline phosphatase, as well as levels of bilirubin, urea and creatinine were revealed by these mice. Postadministration with QC remarkably alleviated the over production of MDA and improved GSH levels in PCM-intoxicated mice blood. In addition, antioxidant enzymes; glutathione peroxidase, glutathione-S-transferase, superoxide dismutase and catalase activities were also improved significantly in these mice. QC had also considerably ameliorated the altered biochemical parameters toward normalcy. Thus, it can be concluded that QC may constitute a remedy against PCM-induced oxidative stress and reno-hepatic injuries.

13C saturation transfer effect of carbon dioxide-bicarbonate exchange catalyzed by carbonic anhydrase in vivo.

Carbonic anhydrase catalyzes reversible hydration of carbon dioxide and dehydration of bicarbonate. In this article we report that the rapid exchange catalyzed by carbonic anhydrase causes a large magnetization (saturation) transfer effect on the 13C signal of bicarbonate at 160.7 ppm in vivo when the resonance of the undetectable carbon dioxide at 125.0 ppm is irradiated with RF pulses. In isoflurane-anesthetized adult rat brain the unidirectional, pseudo first-order rate constant of this exchange in the dehydration direction was determined to be 0.47 +/- 0.05 sec(-1) following intravenous infusion of uniformly 13C-labeled glucose for labeling bicarbonate. Intralateral ventricular administration of the highly specific carbonic anhydrase inhibitor acetazolamide, which is a drug used for treating glaucoma and epilepsy, was also shown to significantly attenuate the observed 13C magnetization transfer effect of the carbon dioxide-bicarbonate exchange in the rat brain.