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Summary: SynNotch receptor technology is a versatile tool that uses the regulatory notch core portion with an extracellular scFv and an intracellular transcription factor that enables to program customized input and output functions in mammalian cells. In this study, we designed a novel synNotch receptor comprising scFv against HBs antigen linked with an intracellular artificial transcription factor and exploited it for viral sensing and cellular immunotherapy. The synNotch receptor expressing cells sensed HBV particles and membrane-bound HBs antigens and responded by expressing reporter molecules, secNL or GFP. We also programmed these cells to dispense antiviral responses such as type I interferon and anti-HBV neutralizing mouse-human chimeric antibodies. Our data reveal that synNotch receptor signaling works for membrane-bound ligands such as enveloped viral particles and proteins borne on liposomal vesicles. This study establishes the concepts of “engineered immunity” where the synNotch platform is utilized for cellular immunotherapy against viral infections. : Immunity; Viral Microbiology; Bioengineering Subject Areas: Immunity, Viral Microbiology, Bioengineering

Cysteine proteases play a crucial role in the development of the human malaria parasites Plasmodium falciparum and Plasmodium vivax. Our earlier studies demonstrated that these enzymes are equipped with specific domains for defined functions and further su

An investigation of the jet trajectories and mixing behavior of venturi-jet mixers, in which turbulent jet injects fluid at an arbitrary angle to mix incompressible fluids, is described in this paper Numerical results of an incompressible cross flow-jet mixing in venturi-jet mixers are presented and validated against experimental results. Jet trajectories, concentration decay of tracer jet, spatial unmixedness, and mixing length are computed for a range of injection angles (45 deg [less than or equal to] [[theta].sub.0] [less than or equal to] 135 deg). Twenty-five cases are studied with five different initial injection angles, each with five different jet momentum ratios. As an example of practical insights that can be gained from such detailed analysis, the resulting flow field is used to obtain an equation for trajectory and tracer concentration in the mixer. The penetration of jet scales with the third root of the jet-to-mainstream momentum ratio and that of with square root of downstream distance. The decay of mean concentration scales with the inverse of axial distance and with the negative root of injection angle. The results show a consistency in the experimental data and simulation has provided a good insight into the flow details and has paved the way in optimization of the geometry based on jet injection angle to get a good mixing efficiency. [DOI: 10.1115/1.4002575] Keywords: jet trajectories, momentum ratio, entrainment, incompressible flow, venturi-jet mixer

Background: Peninsular India, being completely under the influence of monsoonal climate, suffers crop yield variability due to rainfall distribution-induced soil moisture constraints. Timely and appropriate assessment of this rainfall and soil moisture-induced crop yield variability serves as a key for exemplary relief assistance. Per cent available soil moisture (PASM) is one among several drought declaration indices followed by stakeholders in India for declaration of drought, needs re-evaluation as the existing criteria in unable to capture the yield loss due to ineffective classification of PASM categories. This study attempts to revise the agricultural drought classes by PASM based on relationships established between yield of major rainfed crops of the study region and PASM.Methods: Analysis of yield variability due to PASM was carried out based on long term observations in experiments conducted at five dry farming locations (Akola, Parbhani, Kovilpatti, Ananthapuramu and Bengaluru) of peninsular India. The average yield for each category of PASM was calculated and tabulated for regression analysis. The PASM versus yield in each group was correlated and regression equations were developed if significant positive correlations were established.Results: The range of available soil moisture to obtain at least 50 percent of optimum yield in cereals (maize: 26 and finger millet: 52.9 PASM), pulses (pigeon pea: 37.2 PASM), oilseeds (soybean: 26.8 to 30.5, groundnut: 53.8 to 61.7 PASM) and commercial crops (cotton: 26.3 PASM) was 26–61 percent.Conclusion: The revised PASM-based drought classes (0–50 severe; 51–75 mild and 76–100 no drought) would help in drought declaration and precise identification of drought-hit areas for meaningful relief assistance. However, there is further investigation is needed to include a soil component for further fine-tuning of the criteria.

The Pfizer-BioNTech vaccine was one of the essential tools in curtailing the COVID-19 pandemic. Unlike conventional vaccines, this newly approved mRNA vaccine is taken up by cells, which leads to the synthesis of the specific viral Spike antigen. The vaccine was initially introduced for adults, and the immunological profile of adult recipients is well-characterized. The vaccine was approved for paediatric use much later after its efficacy and safety had been confirmed in children. However, the complete picture of how the paediatric immune system in children reacts to the vaccine is not well documented. Therefore, in order to better understand the immune response in children, we analysed the humoral response, immune cell count, and interferon signalling in paediatric vaccine recipients ranging between 5 and 17 years of age. Our findings suggest that the paediatric recipients elicit a robust humoral response that is sustained for at least three months. We also found that the vaccine triggered a transient lymphocytopenia similar to that observed during viral infection. Interestingly, we also found that the vaccine may sensitise the interferon signalling pathway, priming the cells to mount a potent response when exposed to interferons during a subsequent infection. The study offers new insights into the workings of the paediatric immune system and innate immunity, thereby opening the doors for further research in this field.

The study highlights the importance of integrating organic resources, such as vermicompost and biofertilizers with inorganic fertilizers to sustain coconut yields and manage costs. The experimental trial was conducted from 2016 to 2020 in a 47-year-old East Coast Tall coconut garden at Coconut Research Station in South India. The research evaluated four nutrient management treatments viz., T1 (75% Recommended Dose of Fertilizer (RDF) + 25% N organic), T2 (50% RDF + 50% N organic), T3 (100% N organic) and a control (100% RDF). Intercrops included black pepper (Piper nigrum), banana (Musa acuminata) and cocoa (Theobroma cacao). Organic manure significantly improved soil physical properties, water retention and overall soil health. The T2 treatment achieved the highest yields for coconut, cocoa, banana and black pepper. The added soil health parameters supported to these findings, with T2 showing the highest fungal (15.27 × 103 cfu/g of soil) and bacterial populations (17.25 × 105 cfu/g of soil), along with a significant earthworm population (26/m2), indicating enhanced soil ecosystem health. Additionally, soil moisture content was highest under T3 (100% organic) across various depths, followed by T2, highlighting the critical role of organic matter in improving soil moisture conservation. The economic feasibility analysis, including a net present value (NPV), benefit/cost ratio (B/C ratio) and an internal rate of return (IRR), revealed T2 to be the most economically viable nutrient management strategy. This study highlights the economic benefits of intercropping coconuts using an integrated nutrient management (INM) approach, demonstrating its superiority over traditional monocropping practices.

Abstract The COVID-19 pandemic is an unprecedented threat to humanity that has provoked global health concerns. Since the etiopathogenesis of this illness is not fully characterized, the prognostic factors enabling treatment decisions have not been well documented. Accurately predicting the progression of the disease would aid in appropriate patient categorization and thus help determine the best treatment option. Here, we have introduced a proteomic approach utilizing data-independent acquisition mass spectrometry (DIA-MS) to identify the serum proteins that are closely associated with COVID-19 prognosis. Twenty-seven proteins were differentially expressed between severely ill COVID-19 patients with an adverse or favorable prognosis. Ingenuity Pathway Analysis revealed that 15 of the 27 proteins might be regulated by cytokine signaling relevant to interleukin (IL)-1β, IL-6, and tumor necrosis factor (TNF), and their differential expression was implicated in the systemic inflammatory response and in cardiovascular disorders. We further evaluated practical predictors of the clinical prognosis of severe COVID-19 patients. Subsequent ELISA assays revealed that CHI3L1 and IGFALS may serve as highly sensitive prognostic markers. Our findings can help formulate a diagnostic approach for accurately identifying COVID-19 patients with severe disease and for providing appropriate treatment based on their predicted prognosis.

The host innate immune response is the first line of defense against human immunodeficiency virus (HIV) infection. The type I interferon (IFN) response is a robust anti-viral response that induces the transcription of several IFN-stimulated genes (ISGs). However, the effects of ISGs, particularly on the HIV-1 Gag protein, remain largely unknown. Hence, we screened ISG-encoded proteins by bioluminescence resonance energy transfer to identify the crucial host effectors that suppressed Gag function. Consequently, we identified the transmembrane protein MAL as a Gag-interacting ISG product. In fact, ectopic expression of MAL substantially inhibited the production of HIV-1 particles, leading to the translocation, accumulation, and eventual lysosomal degradation of Gag in the host endosomal compartments. Owing to the conserved N-terminal region of MAL, which specifically interacts with HIV-1 Gag, this particular antiviral function of MAL targeting Gag is also conserved among orthologs of various animal species. Notably, the antiviral activity of MAL was partially antagonized by the viral accessory protein Nef, as it interfered with the interaction between MAL and Gag. Therefore, this study reveals a previously unidentified antiviral function of MAL and its viral counteraction. It also sheds new light on therapeutic strategies against HIV-1 infection based on the intrinsic antiviral immunity of host cells.

The successive emergence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants has presented a major challenge in the management of the coronavirus disease (COVID-19) pandemic. There are growing concerns regarding the emerging variants escaping vaccines or therapeutic neutralizing antibodies. In this study, we conducted an epidemiological survey to identify SARS-CoV-2 variants that are sporadically proliferating in vaccine-advanced countries. Subsequently, we created HiBiT-tagged virus-like particles displaying spike proteins derived from the variants to analyze the neutralizing efficacy of the BNT162b2 mRNA vaccine and several therapeutic antibodies. We found that the Mu variant and a derivative of the Delta strain with E484K and N501Y mutations significantly evaded vaccine-elicited neutralizing antibodies. This trend was also observed in the Beta and Gamma variants, although they are currently not prevalent. Although 95.2% of the vaccinees exhibited prominent neutralizing activity against the prototype strain, only 73.8 and 78.6% of the vaccinees exhibited neutralizing activity against the Mu and the Delta derivative variants, respectively. A long-term analysis showed that 88.8% of the vaccinees initially exhibited strong neutralizing activity against the currently circulating Delta strain; the number decreased to 31.6% for the individuals at 6 months after vaccination. Notably, these variants were shown to be resistant to several therapeutic antibodies. Our findings demonstrate the differential neutralization efficacy of the COVID-19 vaccine and monoclonal antibodies against circulating variants, suggesting the need for pandemic alerts and booster vaccinations against the currently prevalent variants.

Summary: The ongoing coronavirus disease 2019 (COVID-19) pandemic is a major global public health concern. Although rapid point-of-care testing for detecting viral antigen is important for management of the outbreak, the current antigen tests are less sensitive than nucleic acid testing. In our current study, we produce monoclonal antibodies (mAbs) that exclusively react with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and exhibit no cross-reactivity with other human coronaviruses, including SARS-CoV. Molecular modeling suggests that the mAbs bind to epitopes present on the exterior surface of the nucleocapsid, making them suitable for detecting SARS-CoV-2 in clinical samples. We further select the optimal pair of anti-SARS-CoV-2 nucleocapsid protein (NP) mAbs using ELISA and then use this mAb pair to develop immunochromatographic assay augmented with silver amplification technology. Our mAbs recognize the variants of concern (501Y.V1-V3) that are currently in circulation. Because of their high performance, the mAbs of this study can serve as good candidates for developing antigen detection kits for COVID-19.

Objective: There is scarce evidence regarding the long-term persistence of neutralizing antibodies among coronavirus disease 2019 (COVID-19) survivors. This study determined neutralizing antibody titers (NT50) and antibodies against spike protein (SP) or nucleocapsid protein (NP) antigens approximately 6 months after the diagnosis of COVID-19.Methods: COVID-19 survivors in Japan were recruited. Serum samples and data related to patients’ characteristics and COVID-19 history were collected. NT50 and titers of antibodies against NP and SP antigens were measured at 20–32 weeks after the first positive severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) test results. Factors associated with NT50 were identified using the multivariable linear regression and the correlations among NT50 and titers of immunoglobulin G (IgG) and total immunoglobulins (Igs) against NP and SP were assessed by Spearman’s correlation.Results: Among 376 participants (median [range] days after testing positive for SARS-CoV-2, 180 (147–224); median [range] years of age, 50 (20–78); 188 [50%] male), most tested positive for NT50 (n = 367, 98%), SP-IgG (n = 344, 91%), SP-total Ig (n = 369, 98%), NP-IgG (n = 314, 84%), and NP-total Ig (n = 365, 97%). Regression analysis indicated that higher BMI, fever, and the requirement of mechanical ventilation or extracorporeal membrane oxygenation were significantly associated with higher NT50. Anti-SP antibodies correlated moderately with NT50 (Spearman’s correlation: 0.63 for SP IgG; 0.57 for SP-total Ig), while the correlation was weak for anti-NP antibodies (0.37 for NP IgG; 0.32 for NP-total Ig).Conclusions: Most COVID-19 survivors had sustained neutralizing antibodies and tested positive for SP-total Ig and NP-total Ig approximately 6 months after infection.

Type-I interferons (IFN-I) are the innate immune system’s principal defense against viral infections. Human immunodeficiency virus-1 (HIV-1) has evolved several ways to suppress or evade the host’s innate immunity in order to survive and replicate to sustain infection. Suppression of IFN-I is one among the multiple escape strategies used by HIV-1 to prevent its clearance. HIV-1 protease which helps in viral maturation has also been observed to cleave host cellular protein kinases. In this study we performed a comprehensive screening of a human kinase library using AlphaScreen assay and identified that TANK binding kinase-1 (TBK1) was cleaved by HIV-1 protease (PR). We demonstrate that PR cleaved TBK1 fails to phosphorylate IFN regulatory factor 3 (IRF3), thereby reducing the IFN-I promoter activity and further reveal that the PR mediated suppression of IFN-I could be counteracted by protease inhibitors (PI) in vitro. We have also revealed that mutations of HIV-1 PR that confer drug resistance to PIs reduce the enzyme’s ability to cleave TBK1. The findings of this study unearth a direct link between HIV-1 PR activity and evasion of innate immunity by the virus, the possible physiological relevance of which warrants to be determined.