Molecular basis of DEL phenotype in the Indian population: Insights from next-generation sequencing analysis of two cases.

The DEL phenotype represents an intriguing and challenging aspect of blood group serology. This condition is characterized by an extremely weak expression of the D antigen on red blood cells, to the extent that it often eludes detection through routine serological methods. Identifying the DEL phenotype necessitates more specialized techniques, such as adsorption and elution tests, to reveal the presence of the D antigen. This distinctive phenotype underscores the complexity and subtlety of blood group genetics and highlights the importance of using advanced methods to accurately classify individuals with this condition, as their ability to form anti-D antibodies can have clinical implications during transfusion and pregnancy scenarios. There is a paucity of data for the DEL phenotype in the Indian population, and the molecular basis has not been elucidated yet. Our investigation delves into the genetic underpinnings of two distinct DEL phenotype cases that pose challenges for resolution through conventional serological techniques. We employ next-generation amplicon sequencing to unravel the intricate genetic landscape underlying these cases. In the D-negative donor, the DEL phenotype was first identified serologically, which was subsequently confirmed by molecular analysis. In the second case, it was associated with an anti-D antibody in a D-positive patient. Initial data analysis unveiled a substantial reduction in coverage across the exonic segments of the RHD gene in both samples, suggesting the potential presence of RHD exon deletions. On both occasions, we identified a homozygous intronic RHD polymorphism that is well established to be linked to the RHD* 01EL.32/RHD*DEL32 variant.

A phase 2 single center open label randomised control trial for convalescent plasma therapy in patients with severe COVID-19.

A single center open label phase 2 randomised control trial (Clinical Trial Registry of India No. CTRI/2020/05/025209) was done to assess clinical and immunological benefits of passive immunization using convalescent plasma therapy. At the Infectious Diseases and Beleghata General Hospital in Kolkata, India, 80 patients hospitalized with severe COVID-19 disease and fulfilling the inclusion criteria (aged more than 18 years, with either mild ARDS having PaO2/FiO2 200-300 or moderate ARDS having PaO2/FiO2 100-200, not on mechanical ventilation) were recruited and randomized into either standard of care (SOC) arm (N = 40) or the convalescent plasma therapy (CPT) arm (N = 40). Primary outcomes were all-cause mortality by day 30 of enrolment and immunological correlates of response to therapy if any, for which plasma abundance of a large panel of cytokines was quantitated before and after intervention to assess the effect of CPT on the systemic hyper-inflammation encountered in these patients. The secondary outcomes were recovery from ARDS and time taken to negative viral RNA PCR as well as to report any adverse reaction to plasma therapy. Transfused convalescent plasma was characterized in terms of its neutralizing antibody content as well as proteome. The trial was completed and it was found that primary outcome of all-cause mortality was not significantly different among severe COVID-19 patients with ARDS randomized to two treatment arms (Mantel-Haenszel Hazard Ratio 0.6731, 95% confidence interval 0.3010-1.505, with a P value of 0.3424 on Mantel-Cox Log-rank test). No adverse effect was reported with CPT. In severe COVID-19 patients with mild or moderate ARDS no significant clinical benefit was registered in this clinical trial with convalescent plasma therapy in terms of prespecified outcomes.

A novel approach to baseline water quality assessment at local and catchment scale: a case study from Berambadi, India.

Optimal design and maintenance are necessary for the sustainability of wastewater treatment systems. In this study, we present the outcome of a novel approach to baseline assessment conducted prior to the design and deployment of a decentralized wastewater treatment system at a school in rural India. The baseline water quality monitoring protocol was deployed to assess (a) the quality and quantity of wastewater (greywater and blackwater) flows from the school and (b) the status of surface water and groundwater quality in the catchment. Hourly greywater flows and water quality trends were monitored across four seasons at the school. Average freshwater consumption at the school was 518 ± 322 L/day for hand washing and 287 ± 97 L/day for cooking meals. Greywater generation showed high hourly variations in COD levels. Greywater generated from hand wash and kitchen sources contributed to 110 g/day and 96 g/day of BOD5 respectively and 214 g/day and 141 g/day of COD respectively. Based on additional data from a self-reporting sanitation survey, the organic contaminant load generated from the toilet was estimated to be 1.5 ± 0.1 kg COD/day. At the catchment scale, both groundwater and surface water quality were monitored seasonally to assess the impact of raw sewage and stormwater inputs. Compared with borewells, high nitrate-N levels (> 10 mg/L) were observed in the village hand pump samples throughout the year. Maximum nitrate-N (16 mg/L) and fecal coliforms (3.9 log MPN/100 mL) levels were observed in surface waters during monsoons, indicating the impact of sewage and surface runoff on water quality. The proposed approach is useful to estimate data on freshwater use and wastewater generation at the school and hence to make the case for, and design of, a sustainable water management intervention.

Nature and Dimensions of Systemic Hyperinflammation and its Attenuation by Convalescent Plasma in Severe COVID-19.

BACKGROUND: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), causing coronavirus disease 2019 (COVID-19), has led to significant morbidity and mortality. While most suffer from mild symptoms, some patients progress to severe disease with acute respiratory distress syndrome (ARDS) and associated systemic hyperinflammation. METHODS: First, to characterize key cytokines and their dynamics in this hyperinflammatory condition, we assessed abundance and correlative expression of a panel of 48 cytokines in patients progressing to ARDS as compared to patients with mild disease. Then, in an ongoing randomized controlled trial of convalescent plasma therapy (CPT), we analyzed rapid effects of CPT on the systemic cytokine dynamics as a correlate for the level of hypoxia experienced by the patients. RESULTS: We identified an anti-inflammatory role of CPT independent of its neutralizing antibody content. CONCLUSIONS: Neutralizing antibodies, as well as reductions in circulating interleukin-6 and interferon-γ-inducible protein 10, contributed to marked rapid reductions in hypoxia in response to CPT. CLINICAL TRIAL REGISTRY OF INDIA: CTRI/2020/05/025209. http://www.ctri.nic.in/.

A prospective study on COVID-19 convalescent plasma donor (CCP) recruitment strategies in a resource constrained blood centre.

Background and Objectives: The COVID-19 pandemic has spread across 87 million people with more than 1·8 million deaths in the world. As there is no definite treatment modality, the use of convalescent plasma has become increasingly popular worldwide. This study aimed to identify an appropriate strategy of donor recruitment and to evaluate the appropriateness of pre-set plasma donation guidelines. Material and Methods: In this prospective study conducted from May to September 2020, the donors were recruited under the following two circumstances: Group I, patients in the post-COVID-19 follow-up in the clinic, and Group II, patients recovered from COVID-19 recruited through mass and electronic media. A pre-set donor selection criteria and laboratory investigation was designed according to national and international guidelines. Approximately 500 ml of COVID-19 convalescent plasma (CCP) was collected from recovered individuals in each group by two different cell separators. The overall donor's attendance rate, deferral rate, adverse events and donor compliance was analysed and compared between the two groups. Results: There was a significant difference in attendance in relation to registration between the groups (P < 0·0001). Donor deferral was significantly higher in group II compared with group I. The single most frequent cause of donor deferral was low antibody index (P = 0·0001). The total donor adverse event rate in CCP donation was significantly lower compared with routine plateletpheresis procedures. The donor's compliance to blood centre's protocol was satisfactory in both the groups. Conclusion: Recruitment of patients in the post-COVID-19 follow-up in the clinic was more effective than the general recruitment through mass and electronic media for convalescence plasma donation in a resource-constrained blood centre.

Provisional green infrastructure: transdisciplinary approaches to address contamination in urban streams.

Surface water contamination has emerged as an area of major concern in rapidly growing cities in the Global South, including and especially in the Indian megacity context. We argue here that nallahs (open drainage channels in Indian megacities) should be more widely recognized as a potential locus of intervention. These combined stormwater and wastewater networks offer opportunities for flexible, frugal and inclusive retrofits to improve surface and groundwater quality. We propose and define the concept of provisional green infrastructure (PGI) as a speculative innovation typology describing in-stream interventions. We argue that PGI should be employed as a shared boundary concept guiding transdisciplinary action and research within the highly unpredictable, space-constrained, and contaminated watersheds. Citing case studies throughout the region and ongoing research in the city of Bangalore, we demonstrate in-stream modifications may be capable of achieving significant improvement in the quality of urban wastewater and may play a complementary role in closing persistent capacity gaps in the operation of both centralized and decentralized treatment practices within megacities. Anticipating the larger diffusion of PGI practices across the region by various early adopters and non-state actors, we suggest a cogent research agenda focused on identifying various generalizable 'upscaling' opportunities for deploying in-stream interventions across various organizational and spatial domains.