Screening Clinical, Laboratory and Host Markers for Diagnosis of Disease Severity in Plasmodium vivax Clinical Samples.

Malaria is one of the most infectious disease that affects lives of million people throughout the world. Recently, there are several reports which indicate Plasmodium vivax (P. vivax) causing severe disease in infected patients from different parts of the world. For P. vivax disease severity, the data related to immunological and inflammatory status in human host is very limited. In the present study clinical parameters, cytokine profile and integrin gene were analyzed in P. vivax clinical patients. A total of 169 P. vivax samples were collected and categorized into severe vivax malaria (SVM; n = 106) and non-severe vivax malaria (NSVM; n = 63) according to WHO severity criteria. We measured host biomarker levels of interferon (IFN-γ), superoxide dismutase (SOD-1), interleukins viz. (IL-6, IL-10), and tumor necrosis factor (TNF-α) in patient plasma samples by ELISA for pro- and anti-inflammatory cytokines in severe malaria. Host integrin gene was genotyped using PCR assay. In our study, thrombocytopenia and anemia were major symptoms in severe P. vivax patients. In analyzed SVM and NSVM groups a significant increase in cytokine levels (IL-10, IL-6, and TNF-α) and anti-oxidant enzyme SOD-1 was found. Our study results also showed a higher pro-inflammatory (TNF-α, IL-6 and IFN-γ) to anti-inflammatory (IL-10) cytokine ratio in severe vivax patients. Integrin gene showed no mutation with respect to thrombocytopenic patients among clinically defined groups. It was observed that severe vivax cases had increased cytokine levels irrespective of age and sex of the patients along with thrombocytopenia and other clinical manifestations. The results of current findings could serve as baseline data for evaluating severe malaria parameters during P. vivax infections and will help in developing an effective biomarker for diagnosis. Supplementary Information: The online version contains supplementary material available at 10.1007/s12088-024-01324-4.

Nationwide spatiotemporal drug resistance genetic profiling from over three decades in Indian Plasmodium falciparum and Plasmodium vivax isolates.

BACKGROUND: Drug resistance is a serious impediment to efficient control and elimination of malaria in endemic areas. METHODS: This study aimed at analysing the genetic profile of molecular drug resistance in Plasmodium falciparum and Plasmodium vivax parasites from India over a ~ 30-year period (1993-2019). Blood samples of P. falciparum and/or P. vivax-infected patients were collected from 14 regions across India. Plasmodial genome was extracted and used for PCR amplification and sequencing of drug resistance genes in P. falciparum (crt, dhps, dhfr, mdr1, k13) and P. vivax (crt-o, dhps, dhfr, mdr1, k12) field isolates. RESULTS: The double mutant pfcrt SVMNT was highly predominant across the country over three decades, with restricted presence of triple mutant CVIET from Maharashtra in 2012. High rates of pfdhfr-pfdhps quadruple mutants were observed with marginal presence of "fully resistant" quintuple mutant ACIRNI-ISGEAA. Also, resistant pfdhfr and pfdhps haplotype has significantly increased in Delhi between 1994 and 2010. For pfmdr1, only 86Y and 184F mutations were present while no pfk13 mutations associated with artemisinin resistance were observed. Regarding P. vivax isolates, the pvcrt-o K10 "AAG" insertion was absent in all samples collected from Delhi in 2017. Pvdhps double mutant SGNAV was found only in Goa samples of year 2008 for the first time. The pvmdr1 908L, 958M and 1076L mutations were highly prevalent in Delhi and Haryana between 2015 and 2019 at complete fixation. One nonsynonymous novel pvk12 polymorphism was identified (K264R) in Goa. CONCLUSIONS: These findings support continuous surveillance and characterization of P. falciparum and P. vivax populations as proxy for effectiveness of anti-malarial drugs in India, especially for independent emergence of artemisinin drug resistance as recently seen in Africa.

First Insight into Drug Resistance Genetic Markers, Glucose-6-Phosphate Dehydrogenase and Phylogenetic Patterns of Misdiagnosed Plasmodium vivax Malaria in Far North Region, Cameroon.

Plasmodium falciparum (Pf) is the predominant malaria species in Africa, but growing rates of non-falciparum species such as P. vivax (Pv) have been reported recently. This study aimed at characterizing drug resistance genes, glucose-6-phosphate dehydrogenase gene (G6PD), and phylogenetic patterns of a Pv + Pf co-infection misdiagnosed as a Pf mono-infection in the Far North region of Cameroon. Only one non-synonymous mutation in the pvdhps gene A383G was found. Pv drug resistance gene sequences were phylogenetically closer to the reference SAL-I strain and isolates from Southeast Asia and Western Pacific countries. Analyzing co-infecting Pf revealed no resistance mutations in Pfmdr1 and Pfk13 genes, but mutations in Pfcrt (C72V73I74E75T76) and Pfdhfr-Pfdhps genes (A16C50I51R59N108L164 - A436A437K540G581S613) were observed. No G6PD deficiency-related mutations were found. This is first study from Cameroon reporting presence of putative drug resistance mutations in Pv infections, especially in the pvdhps gene, and also outlined the absence of a G6PD-deficiency trait in patients.

Characterization of drug resistance genes in Indian Plasmodium falciparum and Plasmodium vivax field isolates.

One of the major challenges for malaria control and elimination is the spread and emergence of antimalarial drug resistance. Mutations in Plasmodium falciparum (Pf) and Plasmodium vivax (Pv) field isolates for five drug resistance genes viz. crt, mdr1, dhps, dhfr and kelch known to confer resistance to choloroquine (CQ), sulfadoxine-pyrimethamine (SP) and artemisinin (ART) and its derivatives were analyzed. A total of 342 symptomatic isolates of P. falciparum (Pf) and P. vivax (Pv) from 1993 to 2014 were retrieved from malaria parasite repository at National Institute of Malaria Research (NIMR). Sample DNA was extracted from dried blood spots and various targeted single nucleotide polymorphisms (SNPs) associated with antimalarial drug resistance were analysed for these isolates. 72S (67.7%) and 76T (83.8%) mutations along with SVMNT haplotype (67.7%) predominated the study population for Pfcrt. The most prevalent SNPs were 108N (73.2%) and 437G (24.8%) and the most prevalent haplotypes were ACNRNI (51.9%) and SAKAA (74.5%) in Pfdhfr and Pfdhps respectively. Only two mutations in Pfmdr1, 86Y (26.31%) and 184F (56.26%), were seen frequently in our study population. No mutations associated with Pfk13 were observed. For Pv, all the studied isolates showed two Pvdhps mutations, 383G and 553G, and two Pfdhfr mutations, 58R and 117N. Similarly, three mutations, viz. 958M, 908L and 1076L were found in Pvmdr1. No variations were observed in Pvcrt-o and Pvk12 genes. Overall, our study demonstrates an increase in mutations associated with SP resistance in both Pf and Pv, however, no single nucleotide polymorphisms (SNPs) associated with ART resistance have been observed for either species. Various SNPs associated with CQ resistance were seen in Pf; whereas only Pvmdr1 associated resistant SNPs were observed in Pv. Therefore, molecular characterization of drug resistance genes is essential for timely monitoring and prevention of malaria by identifying the circulating drug resistant parasites in the country.

Use of Locally Produced Novel Low-Cost 3D Printed Respiratory Muscle Strength Trainer Device (RMSTD) for Long COVID-Rehabilitation: An Innovative Case Report.

Introduction. This article details the development and use of a low-cost, custom RMST device for a patient with long COVID who had received positive airway flow support during ICU treatment. By sharing our successful management of respiratory muscle weakness in a severe COVID-19 patient, we aim to contribute to the broader conversation around effective long COVID management. Case Description. A patient with respiratory muscle weakness used a hospital-made RMST device. The training involved ten short forced exhalations per cycle for five cycles, followed by five long forced exhalations held for five seconds. Additionally, the patient learned lower abdominal and box breathing techniques. The patient showed significant improvement, using the RMST device without supplemental oxygen within 24 hours and completely weaned off by day 5. By discharge, the patient could complete the RMST exercise regime and achieved 290 meters in the 6MWT. After six weeks of outpatient therapy, the patient used the RMST device at 80 cm H2O and increased their 6MWT distance to 390 meters. Device Description. The RMST device was designed for in-house production with a 3D-printed PETG cap, base, and plunger, and a standard pen spring. Its design mimicked a standard PEEP valve with a different geometry. The spring closed the valve until a pressure threshold was reached, allowing airflow. The adjustable pressure threshold ranged from -40 to +40 cm H2O, calibrated in 10 cm H2O increments by measuring displaced water column height. Discussion and Limitation. COVID-19 can cause long-term respiratory issues needing proper management and rehabilitation. Inspiratory muscle training benefits those recovering from severe COVID-19 who were weaned from mechanical ventilation. However, the device's design and production method are unsuitable for large-scale and commercial production. Rehabilitation centers should prepare for postintensive care syndrome in post-COVID-19 individuals, with interprofessional teams addressing various recovery aspects. Early medical attention and therapy can improve patients' quality of life.

Assessment of the immunosuppressive and hemolytic activities of an edible fern, Diplazium esculentum.

CONTEXT: Diplazium esculentum is the most commonly consumed fern throughout Asia and Oceania. Systemic toxicity and pathological effects on its consumption have already been demonstrated. But, the immunosuppressive and hemolytic activities of the boiled Diplazium esculentum (BDE), the form in which human consumes it, have not yet been studied. OBJECTIVE: To investigate the immunosuppressive as well as hemolytic activities, if any, of BDE in Swiss albino mice. MATERIALS AND METHODS: Body weight, relative spleen weight, plaque forming cell assay, hemaggutination antibody (HA) titer assay and macrophage counting were performed in BDE treated mice and respective control groups within a span of 180 days, and in vitro assays such as counting of cultured splenocytes, splenocytes proliferation assay and hemolytic assay were performed to justify the immunomodulatory as well as hemolytic activities of D. esculentum. RESULTS: Body weight and relative spleen weight were significantly decreased in BDE fed mice. Significant decreases were observed in the number of plaques formed, HA titer value and in the number of peritoneal macrophages within a span of 180 d. Significant dose-dependent decrease was observed in the number of cultured splenocytes. Significant dose-dependent increases in the percentage inhibition of splenocyte proliferation as well as percentage of hemolysis were evident by in vitro assays. DISCUSSION: These results suggest that the intake of D. esculentum may evoke immune dysfunction as well as may cause destruction of erythrocytes even after cooking. CONCLUSION: Therefore, the consumption of D. esculentum is alarming and may act as immunosuppressive agent.

The spectrum of clinical biomarkers in severe malaria and new avenues for exploration.

Globally, malaria is a public health concern, with severe malaria (SM) contributing a major share of the disease burden in malaria endemic countries. In this context, identification and validation of SM biomarkers are essential in clinical practice. Some biomarkers (C-reactive protein, angiopoietin 2, angiopoietin-2/1 ratio, platelet count, histidine-rich protein 2) have yielded interesting results in the prognosis of Plasmodium falciparum severe malaria, but for severe P. vivax and P. knowlesi malaria, similar evidence is missing. The validation of these biomarkers is hindered by several factors such as low sample size, paucity of evidence-evaluating studies, suboptimal values of sensitivity/specificity, poor clinical practicality of measurement methods, mixed Plasmodium infections, and good clinical value of the biomarkers for concurrent infections (pneumonia and current COVID-19 pandemic). Most of these biomarkers are non-specific to pathogens as they are related to host response and hence should be regarded as prognostic/predictive biomarkers that complement but do not replace pathogen biomarkers for clinical evaluation of SM patients. This review highlights the importance of research on diagnostic/predictive/therapeutic biomarkers, neglected malaria species, and clinical practicality of measurement methods in future studies. Finally, the importance of omics technologies for faster identification/validation of SM biomarkers is also included.