Seasonal influence of physicochemical and climatic parameters on phytoplankton diversity and abundance pattern in community managed semi-impacted floodplain wetland.

Wetlands provide numerous ecological services and are key habitats for aquatic flora and fauna. In the Beledanga wetland, the current study was conducted for 3 years, from July 2019 to June 2021, to evaluate the seasonal influence of physicochemical parameters on phytoplankton diversity and abundance patterns. Overall 48 genera of phytoplankton were observed. Bacillariophyceae (27%) contributed the maximum to the total phytoplankton density. The total abundance of phytoplankton was found utmost during monsoon (4.081 × 103 unit l-1) and least during post-monsoon (3.316 × 103 unit l-1). One-way analysis of variance indicated significant seasonal differences (p < 0.05) for some genera. The study gave the idea about the most influencing physic-chemical parameters (dissolved oxygen, turbidity, total hardness, Ca2+, and total nitrogen) on the growth of phytoplankton with the help of different multivariate and univariate analysis (canonical correspondence analysis and Karl Pearson's correlation). The study again highlighted that climate parameters (temperature and rainfall) had some effect on the phytoplanktonic groups. Our study conceded that N:P in the studied wetland was less than the Redfield ratio (16:1) in all three seasons, while the Si:P ratio was noticed in the high range (15:1) during pre-monsoon. The value of the Shannon diversity index and Margalef's species richness index were noticed to be > 3, which signified quite rich in phytoplankton diversity. But the value of Algal Pollution Index, which describes the ecological pollution level based on the present algal genera was observed high throughout all seasons, indicating organic load. So in future the studied wetland may get adversely affected with influence of anthropogenic activities. Therefore, for sustainable biodiversity of the waterbody, the anthropogenic activities (retting and intensification of agricultural farming) and macrophytes need to be controlled and regulated.

Role of lactoyl-glutathione lyase of Salmonella in the colonization of plants under salinity stress.

Salmonella, a foodborne human pathogen, can colonize the members of the kingdom Plantae. However, the basis of the persistence of Salmonella in plants is largely unknown. Plants encounter various biotic and abiotic stress agents in soil. We conjectured that methylglyoxal (MG), one of the common metabolites that accumulate in plants during both biotic and abiotic stress, plays a role in regulating the plant-Salmonella interaction. The interaction of Salmonella Typhimurium with plants under salinity stress was investigated. It was observed that wild-type Salmonella Typhimurium can efficiently colonize the root, but mutant bacteria lacking MG detoxifying enzyme, lactoyl-glutathione lyase (Lgl), showed lower colonization in roots exclusively under salinity stress. This colonization defect is due to the poor viability of the mutated bacterial strains under these conditions. This is the first report to prove the role of MG-detoxification genes in the colonization of stressed plants and highlights the possible involvement of metabolic genes in the evolution of the plant-associated life of Salmonella.

Integration of pen aquaculture into ecosystem-based enhancement of small-scale fisheries in a macrophyte dominated floodplain wetland of India.

The rapid degradation, overexploitation, and encroachment of floodplain wetlands have led to considerable decline in fish diversity and production from these invaluable aquatic resources threatening livelihood of the dependent fishers. The climate change evident in the fast few decades has further aggravated the problem of eutrophication causing water stress and sedimentation leading to rampant macrophyte proliferation affecting ecological and economic functioning of these ecosystems. Macrophyte control and management needs serious attention for sustaining ecosystem services provided by these resources. In this direction, pen culture of grass carp Ctenopharyngodon idella as a biocontrol for macrophytes along with Indian major carps was implemented in a co-management mode in Beledanga, a typical floodplain wetland, a gradually shrinking, macrophyte dominated floodplain wetland in lower Ganga basin. Indian major carps Labeo catla (6.28±0.23g), Labeo rohita (5.1±0.12g), Cirrhinus mrigala (3.5±0.08g) were stocked in the ratio 4:3:3 at the rate of 20 Nos.m-2 in pens (0.1ha each) in triplicate. Grass carp (7.1±0.42g) was stocked in pen at the rate of 20Nos.m-2 in duplicate. The fishes were fed with pelleted feed twice a day at the rate of 2-3% of body weight. The seed was overwintered in pens for a period of 90 days from November 2019 to January 2020. Average weight recorded at the end of culture period was 25.13±1.70g, 18.11±0.63g, 14.53±0.87g, and 39.20±1.90g in L. catla, L. rohita, C. mrigala, and C. idella, respectively. The survival of fish ranged from 70 to 81%. Growth performance and feed utilization efficiency of grass carp were significantly higher (p<0.05) compared to other carp species. The pen culture was found to be economically viable with a benefit cost ratio of 1.53. The fishes produced were released back into the open wetland as an additional input for culture-based fisheries. The intervention along with niche-based enhanced stocking led to 24% increase in the fish production from the wetland with grass carp contributing 20-22% of the total catch with 32% increase in revenue generated by the sale of fish within a short span of 1 year. The study successfully demonstrated technological suitability and economic feasibility of pen culture in this wetland and role of grass carp as a potential biocontrol species for macrophyte management. Grass carp stocked in open wetland grew to 0.8 to 1kg within 6 months and 2-2.3kg within a year and could utilize 40-45% of the submerged and emergent macrophytes. Integration and optimization of grass carp will not only aid in habitat management of macrophyte-choked wetlands but will also boost their small-scale fisheries by converting standing macrophyte biomass into protein-rich fish biomass. The enhanced production will also cater to nutritional and livelihood security of the dependent fishers.

Spontaneous Air-leak Syndrome and COVID-19: A Multifaceted Challenge.

Spontaneous air-leak syndromes have emerged as rare but significant complication of Coronavirus disease-2019 (COVID-19) pneumonia in the last few months. This complication has been documented in both spontaneous and mechanically ventilated patients. Although few studies have used computed tomographic scans to confirm the diagnosis, this could be challenging in resource-limited setup. We present a series of 15 cases that highlight the clinical heterogeneity with respect to stage of illness, ventilatory status, and varied clinical scenarios at the time of development of these syndromes. All cases in our series were diagnosed clinically and confirmed by bedside chest X-ray and were managed promptly. Though mortality was not so infrequent in our experience, these air-leak syndromes were not directly attributed as cause of death in these patients. Therefore, high level of clinical suspicion and vigilance is necessary to identify and manage cases of air-leak syndrome. How to cite this article: Sabharwal P, Chakraborty S, Tyagi N, Kumar R, Taneja A. Spontaneous Air-leak Syndrome and COVID-19: A Multifaceted Challenge. Indian J Crit Care Med 2021;25(5):584-587.

Acute Flaccid Quadriparesis in a Recovering COVID-19 Patient: A Clinical Dilemma.

How to cite this article: Sabharwal P, Chakraborty S, Tyagi N, Kumar A. Acute Flaccid Quadriparesis in a Recovering COVID-19 Patient: A Clinical Dilemma. Indian J Crit Care Med 2021;25(2):238-239.

Loss of Fas signaling in fibroblasts impairs homeostatic fibrosis resolution and promotes persistent pulmonary fibrosis.

Idiopathic pulmonary fibrosis (IPF) is a progressive, irreversible fibrotic disease of the distal lung alveoli that culminates in respiratory failure and reduced lifespan. Unlike normal lung repair in response to injury, IPF is associated with the accumulation and persistence of fibroblasts and myofibroblasts, as well as continued production of collagen and other extracellular matrix (ECM) components. Prior in vitro studies have led to the hypothesis that the development of resistance to Fas-induced apoptosis by lung fibroblasts and myofibroblasts contributes to their accumulation in the distal lung tissues of IPF patients. Here, we test this hypothesis in vivo in the resolving model of bleomycin-induced pulmonary fibrosis in mice. Using genetic loss-of-function approaches to inhibit Fas signaling in fibroblasts, potentially novel flow cytometry strategies to quantify lung fibroblast subsets, and transcriptional profiling of lung fibroblasts by bulk and single cell RNA sequencing, we show that Fas is necessary for lung fibroblast apoptosis during homeostatic resolution of bleomycin-induced pulmonary fibrosis in vivo. Furthermore, we show that loss of Fas signaling leads to the persistence and continued profibrotic functions of lung fibroblasts. Our studies provide insights into the mechanisms that contribute to fibroblast survival, persistence, and continued ECM deposition in the context of IPF and how failure to undergo Fas-induced apoptosis impairs fibrosis resolution.

Evaluation of retrograde intubation with different doses of dexmedetomidine infusion: A randomised controlled trial.

BACKGROUND: Retrograde intubation is one of the well-described and alternative methods of difficult airway management. It requires effective sedation and patient preparation. Study was done to evaluate intubating conditions during retrograde guided intubation with two different doses of dexmedetomidine. METHODS: This prospective randomized double blind parallel group trial was planned on 60 patients with difficult airway. Patients were divided in two groups to receive either dexmedetomidine 1.0 µg/kg (Group A) or dexmedetomidine 1.5 µg/kg (Group B) by intravenous (IV) route. The Modified Observer Assessment Awareness and Sedation (OAA/S) was measured as primary outcome and ease of intubation, facial grimace score, cough severity, hemodynamic response, patient recall and discomfort were assessed as secondary outcome during awake retrograde intubation. RESULTS: Groups were comparable in terms of demographic and baseline parameters. OAA/S (P = 0.001), cough severity (P < 0.001), facial grimace score (P < 0.001), grading of discomfort during procedure (P < 0.001) and recall of procedure scale (P = 0.038) were found significantly better/lower in Group B as compared to Group A. Hemodynamic parameters were better in Group B and showed significant difference during the retrograde intubation. However, ease of intubation scale, intubating time and complications were not significantly different (P > 0.05) between the two groups. CONCLUSION: Retrograde intubation can be easily learned and performed with minimal complications. Dexmedetomidine in a dose of 1.5 µg/kg IV is optimum and safe for retrograde intubation with clinically manageable side effects.

Glycolytic reprograming in Salmonella counters NOX2-mediated dissipation of ΔpH.

The microbial adaptations to the respiratory burst remain poorly understood, and establishing how the NADPH oxidase (NOX2) kills microbes has proven elusive. Here we demonstrate that NOX2 collapses the ΔpH of intracellular Salmonella Typhimurium. The depolarization experienced by Salmonella undergoing oxidative stress impairs folding of periplasmic proteins. Depolarization in respiring Salmonella mediates intense bactericidal activity of reactive oxygen species (ROS). Salmonella adapts to the challenges oxidative stress imposes on membrane bioenergetics by shifting redox balance to glycolysis and fermentation, thereby diminishing electron flow through the membrane, meeting energetic requirements and anaplerotically generating tricarboxylic acid intermediates. By diverting electrons away from the respiratory chain, glycolysis also enables thiol/disulfide exchange-mediated folding of bacterial cell envelope proteins during periods of oxidative stress. Thus, primordial metabolic pathways, already present in bacteria before aerobic respiration evolved, offer a solution to the stress ROS exert on molecular targets at the bacterial cell envelope.

Zinc-dependent substrate-level phosphorylation powers Salmonella growth under nitrosative stress of the innate host response.

The metabolic processes that enable the replication of intracellular Salmonella under nitrosative stress conditions engendered in the innate response of macrophages are poorly understood. A screen of Salmonella transposon mutants identified the ABC-type high-affinity zinc uptake system ZnuABC as a critical determinant of the adaptation of Salmonella to the nitrosative stress generated by the enzymatic activity of inducible nitric oxide (NO) synthase of mononuclear phagocytic cells. NO limits the virulence of a znuB mutant in an acute murine model of salmonellosis. The ZnuABC transporter is crucial for the glycolytic function of fructose bisphosphate aldolase, thereby fueling growth of Salmonella during nitrosative stress produced in the innate response of macrophages. Our investigations demonstrate that glycolysis mediates resistance of Salmonella to the antimicrobial activity of NO produced in an acute model of infection. The ATP synthesized by substrate-level phosphorylation at the payoff phase of glycolysis and acetate fermentation powers the replication of Salmonella experiencing high levels of nitrosative stress. In contrast, despite its high potential for ATP synthesis, oxidative phosphorylation is a major target of inhibition by NO and contributes little to the antinitrosative defenses of intracellular Salmonella. Our investigations have uncovered a previously unsuspected conjunction between zinc homeostasis, glucose metabolism and cellular energetics in the adaptation of intracellular Salmonella to the reactive nitrogen species synthesized in the innate host response.

Lactoylglutathione lyase, a critical enzyme in methylglyoxal detoxification, contributes to survival of Salmonella in the nutrient rich environment.

Glyoxalase I which is synonymously known as lactoylglutathione lyase is a critical enzyme in methylglyoxal (MG) detoxification. We assessed the STM3117 encoded lactoylglutathione lyase (Lgl) of Salmonella Typhimurium, which is known to function as a virulence factor, due in part to its ability to detoxify methylglyoxal. We found that STM3117 encoded Lgl isomerises the hemithioacetal adduct of MG and glutathione (GSH) into S-lactoylglutathione. Lgl was observed to be an outer membrane bound protein with maximum expression at the exponential growth phase. The deletion mutant of S. Typhimurium (Δlgl) exhibited a notable growth inhibition coupled with oxidative DNA damage and membrane disruptions, in accordance with the growth arrest phenomenon associated with typical glyoxalase I deletion. However, growth in glucose minimal medium did not result in any inhibition. Endogenous expression of recombinant Lgl in serovar Typhi led to an increased resistance and growth in presence of external MG. Being a metalloprotein, Lgl was found to get activated maximally by Co(2+) ion followed by Ni(2+), while Zn(2+) did not activate the enzyme and this could be attributed to the geometry of the particular protein-metal complex attained in the catalytically active state. Our results offer an insight on the pivotal role of the virulence associated and horizontally acquired STM3117 gene in non-typhoidal serovars with direct correlation of its activity in lending survival advantage to Salmonella spp.

Cells producing their own nemesis: understanding methylglyoxal metabolism.

Methylglyoxal, which is technically known as 2-oxopropanal or pyruvaldehyde, shows typical reactions of carbonyl compounds as it has both an aldehyde and a ketone functional group. It is an extremely cytotoxic physiological metabolite, which is generated by both enzymatic and nonenzymatic reactions. The deleterious nature of the compound is due to its ability to glycate and crosslink macromolecules like protein and DNA, respectively. However, despite having toxic effects on cellular processes, methylglyoxal retains its efficacy as an anticancer drug. Indeed, methylglyoxal is one of the well-known anticancer therapeutic agents used in the treatment. Several studies on methylglyoxal biology revolve around the manifestations of its inhibitory effects and toxicity in microbial growth and diabetic complications, respectively. Here, we have revisited the chronology of methylglyoxal research with emphasis on metabolism of methylglyoxal and implications of methylglyoxal production or detoxification on bacterial pathogenesis and disease progression.

Salmonella methylglyoxal detoxification by STM3117-encoded lactoylglutathione lyase affects virulence in coordination with Salmonella pathogenicity island 2 and phagosomal acidification.

Intracellular pathogens such as Salmonella enterica serovar Typhimurium (S. Typhimurium) manipulate their host cells through the interplay of various virulence factors. A multitude of such virulence factors are encoded on the genome of S. Typhimurium and are usually organized in pathogenicity islands. The virulence-associated genomic stretch of STM3117-3120 has structural features of pathogenicity islands and is present exclusively in non-typhoidal serovars of Salmonella. It encodes metabolic enzymes predicted to be involved in methylglyoxal metabolism. STM3117-encoded lactoylglutathione lyase significantly impacts the proliferation of intracellular Salmonella. The deletion mutant of STM3117 (Δlgl) fails to grow in epithelial cells but hyper-replicates in macrophages. This difference in proliferation outcome was the consequence of failure to detoxify methylglyoxal by Δlgl, which was also reflected in the form of oxidative DNA damage and upregulation of kefB in the mutant. Within macrophages, the toxicity of methylglyoxal adducts elicits the potassium efflux channel (KefB) in the mutant which subsequently modulates the acidification of mutant-containing vacuoles (MCVs). The perturbation in the pH of the MCV milieu and bacterial cytosol enhances the Salmonella pathogenicity island 2 translocation in Δlgl, increasing its net growth within macrophages. In epithelial cells, however, the maturation of Δlgl-containing vacuoles were affected as these non-phagocytic cells maintain less acidic vacuoles compared to those in macrophages. Remarkably, ectopic expression of Toll-like receptors 2 and 4 on epithelial cells partially restored the survival of Δlgl. This study identified a novel metabolic enzyme in S. Typhimurium whose activity during intracellular infection within a given host cell type differentially affected the virulence of the bacteria.

Division of the Salmonella-containing vacuole and depletion of acidic lysosomes in Salmonella-infected host cells are novel strategies of Salmonella enterica to avoid lysosomes.

Salmonella has evolved several strategies to counteract intracellular microbicidal agents like reactive oxygen and nitrogen species. However, it is not yet clear how Salmonella escapes lysosomal degradation. Some studies have demonstrated that Salmonella can inhibit phagolysosomal fusion, whereas other reports have shown that the Salmonella-containing vacuole (SCV) fuses/interacts with lysosomes. Here, we have addressed this issue from a different perspective by investigating if the infected host cell has a sufficient quantity of lysosomes to target Salmonella. Our results suggest that SCVs divide along with Salmonella, resulting in a single bacterium per SCV. As a consequence, the SCV load per cell increases with the division of Salmonella inside the host cell. This demands more investment from the host cell to counteract Salmonella. Interestingly, we observed that Salmonella infection decreases the number of acidic lysosomes inside the host cell both in vitro and in vivo. These events potentially result in a condition in which an infected cell is left with insufficient acidic lysosomes to target the increasing number of SCVs, which favors the survival and proliferation of Salmonella inside the host cell.

lac repressor is an antivirulence factor of Salmonella enterica: its role in the evolution of virulence in Salmonella.

The genus Salmonella includes many pathogens of great medical and veterinary importance. Bacteria belonging to this genus are very closely related to those belonging to the genus Escherichia. lacZYA operon and lacI are present in Escherichia coli, but not in Salmonella enterica. It has been proposed that Salmonella has lost lacZYA operon and lacI during evolution. In this study, we have investigated the physiological and evolutionary significance of the absence of lacI in Salmonella enterica. Using murine model of typhoid fever, we show that the expression of LacI causes a remarkable reduction in the virulence of Salmonella enterica. LacI also suppresses the ability of Salmonella enterica to proliferate inside murine macrophages. Microarray analysis revealed that LacI interferes with the expression of virulence genes of Salmonella pathogenicity island 2. This effect was confirmed by RT-PCR and Western blot analysis. Interestingly, we found that SBG0326 of Salmonella bongori is homologous to lacI of Escherichia coli. Salmonella bongori is the only other species of the genus Salmonella and it lacks the virulence genes of Salmonella pathogenicity island 2. Overall, our results demonstrate that LacI is an antivirulence factor of Salmonella enterica and suggest that absence of lacI has facilitated the acquisition of virulence genes of Salmonella pathogenicity island 2 in Salmonella enterica making it a successful systemic pathogen.