Phase 2 randomised placebo-controlled trial of spironolactone and dexamethasone versus dexamethasone in COVID-19 hospitalised patients in Delhi.

BACKGROUND: In this phase 2 randomised placebo-controlled clinical trial in patients with COVID-19, we hypothesised that blocking mineralocorticoid receptors using a combination of dexamethasone to suppress cortisol secretion and spironolactone is safe and may reduce illness severity. METHODS: Hospitalised patients with confirmed COVID-19 were randomly allocated to low dose oral spironolactone (50 mg day 1, then 25 mg once daily for 21 days) or standard of care in a 2:1 ratio. Both groups received dexamethasone 6 mg daily for 10 days. Group allocation was blinded to the patient and research team. Primary outcomes were time to recovery, defined as the number of days until patients achieved WHO Ordinal Scale (OS) category ≤ 3, and the effect of spironolactone on aldosterone, D-dimer, angiotensin II and Von Willebrand Factor (VWF). RESULTS: One hundred twenty patients with PCR confirmed COVID were recruited in Delhi from 01 February to 30 April 2021. 74 were randomly assigned to spironolactone and dexamethasone (SpiroDex), and 46 to dexamethasone alone (Dex). There was no significant difference in the time to recovery between SpiroDex and Dex groups (SpiroDex median 4.5 days, Dex median 5.5 days, p = 0.055). SpiroDex patients had significantly lower D-dimer levels on days 4 and 7 (day 7 mean D-dimer: SpiroDex 1.15 µg/mL, Dex 3.15 µg/mL, p = 0.0004) and aldosterone at day 7 (SpiroDex 6.8 ng/dL, Dex 14.52 ng/dL, p = 0.0075). There was no difference in VWF or angiotensin II levels between groups. For secondary outcomes, SpiroDex patients had a significantly greater number of oxygen free days and reached oxygen freedom sooner than the Dex group. Cough scores were no different during the acute illness, however the SpiroDex group had lower scores at day 28. There was no difference in corticosteroid levels between groups. There was no increase in adverse events in patients receiving SpiroDex. CONCLUSION: Low dose oral spironolactone in addition to dexamethasone was safe and reduced D-dimer and aldosterone. Time to recovery was not significantly reduced. Phase 3 randomised controlled trials with spironolactone and dexamethasone should be considered. TRIAL REGISTRATION: The trial was registered on the Clinical Trials Registry of India TRI: CTRI/2021/03/031721, reference: REF/2021/03/041472. Registered on 04/03/2021.

Root system architecture in rice: impacts of genes, phytohormones and root microbiota.

To feed the continuously expanding world's population, new crop varieties have been generated, which significantly contribute to the world's food security. However, the growth of these improved plant varieties relies primarily on synthetic fertilizers, which negatively affect the environment and human health; therefore, continuous improvement is needed for sustainable agriculture. Several plants, including cereal crops, have the adaptive capability to combat adverse environmental changes by altering physiological and molecular mechanisms and modifying their root system to improve nutrient uptake efficiency. These plants operate distinct pathways at various developmental stages to optimally establish their root system. These processes include changes in the expression profile of genes, changes in phytohormone level, and microbiome-induced root system architecture (RSA) modification. Several studies have been performed to understand microbial colonization and their involvement in RSA improvement through changes in phytohormone and transcriptomic levels. This review highlights the impact of genes, phytohormones, and particularly root microbiota in influencing RSA and provides new insights resulting from recent studies on rice root as a model system and summarizes the current knowledge about biochemical and central molecular mechanisms.

CC-type glutaredoxin, OsGrx_C7 plays a crucial role in enhancing protection against salt stress in rice.

Soil salinity is one of the critical issue worldwide that adversely affect soil fertility. Salt stress significantly limits crop yield and grain quality; therefore, there is an urgent need to develop a strategy to improve salt stress tolerance. In present study, we reported that rice glutaredoxin (OsGrx_C7) plays a positive response in salt induced stress. Gene expression analysis, silencing, and overexpression of OsGrx_C7 gene were used to discover the role of OsGrx_C7 in response to salt stress. Gene expression analysis suggested that OsGrx_C7 expression was induced under salt stress and ubiquitously expressed in rice including root and shoot. The silencing of osgrx_c7 gene leads to increased sensitivity to salt stress, indicating its importance in salt stress tolerance. A gain-of-function approach showed that OsGrx_C7 may act as an important determinant in salt stress, compared with WT, and revealed higher biomass accumulation, improved root and plant growth under salt stress. Under salt stress condition, OsGrx_C7 overexpressing rice plants showed lower level of lipid peroxidation and Na+/K+ ratio, while proline accumulation, soluble sugar content and GSH/GSSG ratio was higher compared to WT. Furthermore, expression analysis suggested that OsGrx_C7 acted as positive regulator of salt tolerance by reinforcing the expression of transporters (OsHKT2;1, OsHKT1;5 and OsSOS1) engaged in Na+ homeostasis in overexpressing plants. Overall our study revealed that OsGrx_C7 emerged as a key mediator in response to salt stress in rice and could be used for engineering tolerance against salt stress in rice and other crops.

Growth, physiological and proteomic responses in field grown wheat varieties exposed to elevated CO2 under high ambient ozone.

The present study investigated growth, biochemical, physiological, yield and proteomic changes in 3 wheat varieties exposed to elevated CO2 (515 ppm) in a background of high ambient ozone in field. Ethylenediurea (EDU) was used as antiozonant. Average ozone concentration was 59 ppb and was sufficient enough to exert phytotoxic effects. Elevated carbon dioxide (eCO2) and EDU application individually or in combination negated the adverse effects of ozone by modulating antioxidants and antioxidative enzymes. Differential leaf proteomics revealed that at vegetative stage major changes in protein abundance were due to EDU treatment (47, 52 and 41 proteins in PBW-343, LOK1 and HD-2967, respectively). Combined treatment of eCO2 and EDU was more responsible for changes in 37 proteins during flowering stage of PBW-343 and LOK1. Functional categorization revealed more than 60% differentially abundant protein collectively belonging to carbon metabolism, protein synthesis assembly and degradation and photosynthesis. At both the growth stages, LOK1 was more responsive to eCO2 and combined treatment (eCO2 + EDU). HD-2967 was more positively responsive to EDU and combined treatment. eCO2 in combination of EDU protected these varieties against high ambient O3.

Proteomic changes may lead to yield alteration in maize under carbon dioxide enriched condition.

In the present study, the effect of elevated CO2 on growth, physiology, yield and proteome was studied on two maize (Zea mays L.) varieties grown under Free-air CO2 enrichment. Growth in high CO2 (530 ppm) did not affect either photosynthesis or pigment contents in both varieties. Reduced MDA content, antioxidant and antioxidative enzymes levels were observed in both varieties in response to high CO2. PEHM-5 accumulated more biomass than SMH-3031 under eCO2. PEHM-5 also had more seed starch and total soluble sugar than SMH-3031. However, SMH-3031 had increased number of seed per cob than PEHM-5. Interestingly, thousand seed weight was significantly increased in PEHM-5 only, while it was decreased in SMH-3031 under eCO2. We observed increased seed size in PEHM-5, while the size of the SMH-3031 seeds remained unaltered. Leaf proteomics revealed more abundance of proteins related to Calvin cycle, protein synthesis assembly and degradation, defense and redox homeostasis in PEHM-5 that contributed to better growth and yield in elevated CO2. While in SMH-3031 leaf, proteins related to Calvin cycle, defense and redox homeostasis were less abundant in elevated CO2 resulting in average growth and yield. The results showed a differential response of two maize varieties to eCO2.

Adaptive fitness of Sapindus emarginatus Vahl populations towards future climatic regimes and the limiting factors of its distribution.

Sapindus emarginatus Vahl (Sapindaceae) also known as 'Indian Soap nut' is significantly important for saponin content in its fruits. However, its current population in India is heavily fragmented due to a lack of sustainable harvesting practices. Moreover, changing climatic regimes may further limit its distribution and possibly compromise the survival of the species in nature. The aim of the present study was to: predict the future distribution range of S. emarginatus; identify the bioclimatic variables limiting this distribution and to evaluate its adaptive fitness and genomic resilience towards these variables. To determine future species distribution range and identify limiting bioclimatic variables, we applied two different ecological niche models (ENMs; BioClim and MaxEnt) on real occurrence data (n = 88 locations). The adaptive fitness of the species was evaluated by quantifying the genetic variability with AFLP markers and marker-environmental associations, using AFLP-associated Bayesian statistics. We found 77% overlap between the baseline (2030) and predicted (2100) species distribution ranges, which were primarily determined by maximum temperature (TMAX) and mean annual precipitation (MAP). The TMAX and MAP contributed 43.1% and 27.1%, respectively to ENM model prediction. Furthermore, AFLP loci significantly associated with bioclimatic variables, and TMAX and MAP represent the lowest proportion (6.15%), confirming to the severe response of the species genome towards these variables. Nevertheless, the very low Linkage disequilibrium (LD) in these loci (4.54%) suggests that the current sensitivity to TMAX and MAP is subject to change during recombination. Moreover, a combination of high heterozygosity (0.40-0.43) and high within-population variability (91.63 ± 0.31%) confirmed high adaptive fitness to maintain reproductive success. Therefore, the current populations of S. emarginatus have substantial genomic resilience towards future climate change, albeit significant conservation efforts (including mass multiplication) are warranted to avoid future deleterious impacts of inbreeding depression on the fragmented populations.

Comparison of rapid bedside tests for phosphorylated insulin-like growth factor-binding protein 1 and fetal fibronectin to predict preterm birth.

OBJECTIVE: To compare the accuracy of rapid bedside tests for phosphorylated insulin-like growth factor-binding protein 1 (phIGFBP-1) and fetal fibronectin (fFN) to predict preterm delivery among women with threatened preterm labor. METHODS: A prospective observational study was conducted among women with a singleton pregnancy of 28-36 weeks, intact membranes, and symptoms suggestive of preterm labor who attended a center in New Delhi, India, between April 1, 2011, and March 31, 2014. Rapid bedside tests were performed at presentation to measure the levels of phIGFBP-1 and fFN in cervicovaginal secretions. All patients were managed as per the standard hospital protocol and followed up until delivery. RESULTS: Data were available for 468 participants. For delivery before 37 weeks, the phIGFBP-1 test exhibited a sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of 81.1%, 97.1%, 95.2% and 87.7%, respectively. The sensitivity, specificity, PPV, and NPV for the fFN test were 19.4%, 99.4%, 97.4%, and 63.2%, respectively. The phIGFBP-1 test displayed higher sensitivity and NPV than did the fFN test for delivery before 34 weeks and within 7 days of testing (P<0.05 for all). CONCLUSION: The rapid bedside test for phIGFBP-1 was more reliable in the prediction of preterm delivery than was the equivalent test for fFN.

Influence of Sulfur Induced Stress on Oxidative Status and Antioxidative Machinery in Leaves of Allium cepa L.

A pot culture experiment was carried out to assess the effect of sulfur stress on growth, oxidative status, and antioxidative metabolism. Onion plants were treated with three different levels of sulfur, namely, 1.0, 4.0, and 8.0 mM S L(-1). Plants raised with 4.0 mM S L(-1) represent sufficient growth for the best vegetative yield. Plants supplied with 1.0 and 8.0 mM S L(-1) showed retarded growth, chlorosis, and reduction in biomass and photoassimilatory pigments. Tissue sulfur concentration and cysteine were increased with increasing sulfur supply. Carbohydrates (sugars and starch) were accumulated in sulfur stressed plants. Hydrogen peroxide levels were increased in sulfur stressed plants. Thiobarbituric acid reactive substances levels were also increased which was an indicator of lipid peroxidation. Enzymatic (superoxide dismutase, catalase, peroxidase, ascorbate peroxidase, and glutathione reductase) and nonenzymatic (asorbate) antioxidative components were enhanced in sulfur stressed plants. Glutathione was increased with increasing sulfur supply. The present study showed that the adverse effects of inadequate sulfur supply result in irregular metabolic activities and antioxidant machinery.

Foliar application of Zn at flowering stage improves plant's performance, yield and yield attributes of black gram.

Black gram plants subjected to varying levels of Zn supply (0.01 to 10 microM Zn) showed optimum growth and dry matter yield in plants receiving 1 microM Zn. The dry matter yield of plants decreased in plants receiving 0.01 and 0.1 microM Zn (deficient) and excess levels of Zn (2 and 10 microM Zn). The plants grown with Zn deficient supply showed delayed flowering, premature bud abscission, reduced size of anthers, pollen producing capacity, pollen viability and stigma receptivity resulting in poor pod formation and seed yield. Providing Zn as a foliar spray at pre-flowering stage minimized the severity of Zn deficiency on reproductive structure development and enhanced the seed nutritional status by enhancing seed Zn density, seed carbohydrate (sugar and starch content) and storage proteins (albumins, globulins, glutenins, and prolamines).

The impact of foliar boron sprays on reproductive biology and seed quality of black gram.

An experiment was conducted under glass house condition to study the effect of foliar application of boron (B) on reproductive biology and seed quality of black gram (Vigna mungo). Black gram (V. mungo L. var. DPU-88-31) was grown under controlled sand culture condition at deficient and sufficient B levels. After 32 days of sowing B deficient plants were sprayed with three concentrations of B (0.05%, 0.1% and 0.2% borax) at three different stages of reproductive development, i.e. prior to flowering, initiation of bud formation and after bud formation. Deficient B supply decreased the anther and pollen size, pollen tube growth, pollen viability as well as stigmatic receptivity which were increased by foliar B application. Foliar spray at all the three concentrations and at all stages increased the yield parameters like number of pods, pod size and number of seeds formed per plant. Foliar B application also improved the seed yield and seed quality in terms of storage seed proteins (albumin, globulin, glutenin and prolamin) and carbohydrates (sugars and starch) in black gram. The foliar application of B in appropriate doses (particularly 0.1%) after bud formation made quantitative and qualitative improvement in seed yield of black gram by supplementing additional/critical B requirements for reproductive development.

Studies on antioxidative enzymes induced by cadmium in pea plants (Pisum sativum).

Pea plants (Pisum sativum cv. Swati) exposed to different concentration of cadmium (50,100, 200 microM Cd) under controlled glass house conditions were quantified for different physiological parameters and antioxidative enzymes. In pea plants, Cd produced a significant inhibition of growth and induced chlorosis, marginal yellowing and necrosis in young leaves, the effect being most pronounced at 200 microM Cd supply. An alteration in the activated oxygen metabolism of pea plants were also detected as evidenced by an increase in concentration of H2O2 and TBARS along with decrease in the chlorophyll and carotenoid concentration in leaves. Cadmium toxicity induced an increase in non-protein thiol, ascorbate, proline and cysteine concentration. A significant increment in the activity of SOD, APX and GR, and a decrease in CAT was observed as a result of Cd treatment. The enhanced activity of SOD and inhibition of CAT and POD produces a high build up of H2O2 which appears to be the main cause of oxidative stress due to Cd toxicity in pea plants.

Zinc is critically required for pollen function and fertilisation in lentil.

The role of zinc (Zn) in reproduction of lentil (Lens culinaris Medik. cv. DPL 15) and the extent to which the Zn requirement for reproduction can be met through supplementation of Zn at the time of initiation of the reproductive phase have been investigated. Low supply (0.1micromol/L) of Zn reduced the size of anthers, the pollen producing capacity and the size and viability of the pollen grains. Scanning electron microscopy (SEM) of pollen grains of Zn deficient plants showed enhanced thickening of exine and wide and raised muri. In vitro germination of pollen grains was reduced by >50% and growth of pollen tubes was retarded. Unlike Zn sufficient plants, the cuticle around the stigmatic papillae of Zn deficient plants remained intact, preventing the interaction between pollen grains and stigmatic exudates that provides the polarity for the growth of pollen tubes through the stylar tract. Zn deficiency increased the activity of acid phosphatase and peroxidase in extracts of pollen grains. Histochemical localisation on the stigmatic surface and native PAGE of the enzyme extracts of pollen grain and stigma exudates showed enhanced expression of acid phosphatase and peroxidase and suppressed expression of esterase in response to Zn deficiency. Zn deficiency reduced the setting of seeds and also their viability. The effect on seed setting was more marked than on in vitro germination of pollen grains, suggesting that the latter was not the exclusive cause of inhibition of fertility. Possibly, loss of fertility was also caused by impairment in pollen-pistil interaction conducive to pollen tube growth and fertilisation. Impairment in pollen structure and function and seed setting was observed even when plants were deprived of Zn at the time of flowering, but to a lesser extent than in plants maintained with low Zn supply from the beginning. Increasing the Zn supply from deficient to sufficient at the initiation of flowering decreased the severity of Zn deficiency effects on pollen and stigma morphology, pollen fertility and seed yield. In conclusion, structural and functional changes induced in pollen grains and stigma of Zn deficient plants and associated decrease in seed setting of lentil indicate a critical requirement of Zn for pollen function and fertilisation that can be partially met by supplementing Zn at the onset of the reproductive phase.