Genotyping var Gene DBL1α Domain of Severe and Non-severe Plasmodium falciparum Patients.

This study analysed the genetic diversity of DBL1α domain of Plasmodium falciparum var gene in severe and non-severe malaria patients from Delhi and Mewat in Northern India. After confirming P. falciparum infection, samples were cloned and the var gene DBL1α domain was sequenced. Out of 377 cloned DBL sequences, 194 were from severe samples and 183 from non-severe samples. Proportion of DBL1α sequences belonging to groups 1, 4 and 5 were significantly higher in severe isolates as compared to non-severe isolates-group 1 (4.1% vs 1.09%, P = 0.0333), group 4 (69.58% vs 74.31%, P < 0.0001), and group 5 (19.58% vs 10.38%, P < 0.0001). Conversely, higher proportion of group 2 was observed in non-severe isolates (0% vs 3.82%, P = 0.0350). Highest diversity was seen in PoLV4 motif of severe and non-severe isolates and like other DBL1α sequences reported from several geographical areas (Africa, Americas, Asia, and Oceania). A total of 247 DBL1α domain haplotypes were found in this study where 139 (56.27%) haplotypes are novel and not reported from India till date. These findings could aid in developing effective malaria interventions, including vaccine and drug targets, by understanding the existing antigenic diversity and vulnerabilities in the parasite's genetic makeup. Supplementary Information: The online version contains supplementary material available at 10.1007/s12088-024-01200-1.

Genetic diversity and natural selection analysis of VAR2CSA and vir genes: implication for vaccine development.

Variable surface antigens (VSAs) encoded by var and vir genes in Plasmodium falciparum and Plasmodium vivax, respectively, are known to be involved in malaria pathogenesis and host immune escape through antigenic variations. Knowledge of the genetic diversity of these antigens is essential for malaria control and effective vaccine development. In this study, we analysed the genetic diversity and evolutionary patterns of two fragments (DBL2X and DBL3X) of VAR2CSA gene and four vir genes (vir 4, vir 12, vir 21 and vir 27) from different endemic regions, including Southeast Asia and sub-Saharan Africa. High levels of segregating sites (S) and haplotype diversity (Hd) were observed in both var and vir genes. Among vir genes, vir 12 (S = 131, Hd = 0.996) and vir 21 (S = 171, Hd = 892) were found to be more diverse as compared to vir 4 (S = 11, Hd = 0.748) and vir 27 (S = 23, Hd = 0.814). DBL2X (S = 99, Hd = 0.996) and DBL3X (S = 307, Hd = 0.999) fragments showed higher genetic diversity. Our analysis indicates that var and vir genes are highly diverse and follow the similar evolutionary pattern globally. Some codons showed signatures of positive or negative selection pressure, but vir and var genes are likely to be under balancing selection. This study highlights the high variability of var and vir genes and underlines the need of functional experimental studies to determine the most relevant allelic forms for effective progress towards vaccine formulation and testing.

Genome-wide expression analysis of novel heat-responsive microRNAs and their targets in contrasting wheat genotypes at reproductive stage under terminal heat stress.

Introduction: Heat stress at terminal stage of wheat is critical and leads to huge yield losses worldwide. microRNAs (miRNAs) play significant regulatory roles in gene expression associated with abiotic and biotic stress at the post-transcriptional level. Methods: In the present study, we carried out a comparative analysis of miRNAs and their targets in flag leaves as well as developing seeds of heat tolerant (RAJ3765) and heat susceptible (HUW510) wheat genotypes under heat stress and normal conditions using small RNA and degradome sequencing. Results and discussion: A total of 84 conserved miRNAs belonging to 35 miRNA families and 93 novel miRNAs were identified in the 8 libraries. Tae-miR9672a-3p, tae-miR9774, tae-miR9669-5p, and tae-miR5048-5p showed the highest expression under heat stress. Tae-miR9775, tae-miR9662b-3p, tae-miR1120a, tae-miR5084, tae-miR1122a, tae-miR5085, tae-miR1118, tae-miR1130a, tae-miR9678-3p, tae-miR7757-5p, tae-miR9668-5p, tae-miR5050, tae-miR9652-5p, and tae-miR9679-5p were expressed only in the tolerant genotype, indicating their role in heat tolerance. Comparison between heat-treated and control groups revealed that 146 known and 57 novel miRNAs were differentially expressed in the various tissues. Eight degradome libraries sequence identified 457 targets of the differentially expressed miRNAs. Functional analysis of the targets indicated their involvement in photosynthesis, spliceosome, biosynthesis of nucleotide sugars and protein processing in the endoplasmic reticulum, arginine and proline metabolism and endocytosis. Conclusion: This study increases the number of identified and novel miRNAs along with their roles involved in heat stress response in contrasting genotypes at two developing stages of wheat.

Trends in clinical features and severity of Plasmodium vivax malaria among children at tertiary care center in North India.

BACKGROUND: Malaria is a significant cause of morbidity and mortality in adults and children. Plasmodium falciparum is the primary cause of severe malaria, but recently Plasmodium vivax is also recognized to cause severe malaria-associated morbidity and mortality. The study focuses on determining the mortality related to severity parameters in individuals under 12 years and their critical presentation in P.vivax malaria-infected children. METHODS: A prospective cross-sectional hospital-based study was conducted at Safdarjung Hospital, New Delhi, and ICMR-NIMR, New Delhi. All clinically suspected cases were admitted for screening. Exclusion criteria (rapid malaria antigen test, microscopy and medication history) were applied to all the admitted patients (n = 221) to obtain P.vivax patients only. Patients aged ≤ 12 years were included in the study. DNA was extracted from dried blood spots and amplified by nested PCR, followed by visualization on gel electrophoresis. RESULT: A total of 221 clinically suspected cases of malaria were screened for P.vivax. After implementing various exclusion criteria, 45/221 cases were enrolled for the study, among which 44.4% (20/45) of children had the symptoms of severe malaria in terms of cerebral malaria, thrombocytopenia, anemia, pancytopenia, acute respiratory distress syndrome and hemophagocytic lymphohistiocytosis. CONCLUSION: Plasmodium vivax mono-infection can cause severe manifestation and must be treated as P.falciparum without any delay because it may lead to increased morbidity and mortality. A changing trend in clinical symptoms has shown in P.vivax which was an earlier phenomenon of P.falciparum.

IL-13 and the hydroperoxy fatty acid 13(S)HpODE play crucial role in inducing an apoptotic pathway in cancer cells involving MAO-A/ROS/p53/p21 signaling axis.

This study depicted the effect of IL-13 and 13(S)HpODE (the endogenous product during IL-13 activation) in the process of cancer cell apoptosis. We examined the role of both IL-13 and 13(S)HpODE in mediating apoptotic pathway in three different in vitro cellular models namely A549 lung cancer, HCT116 colorectal cancer and CCF52 GBM cells. Our data showed that IL-13 promotes apoptosis of A549 lung carcinoma cells through the involvement of 15-LO, PPARγ and MAO-A. Our observations demonstrated that IL-13/13(S)HpODE stimulate MAO-A-mediated intracellular ROS production and p53 as well as p21 induction which play a crucial role in IL-13-stimulated A549 cell apoptosis. We further showed that 13(S)HpODE promotes apoptosis of HCT116 and CCF52 cells through the up-regulation of p53 and p21 expression. Our data delineated that IL-13 stimulates p53 and p21 induction which is mediated through 15-LO and MAO-A in A549 cells. In addition, we observed that PPARγ plays a vital role in apoptosis as well as in p53 and p21 expression in A549 cells in the presence of IL-13. We validated our observations in case of an in vivo colon cancer tumorigenic study using syngeneic mice model and demonstrated that 13(S)HpODE significantly reduces solid tumor growth through the activation of apoptosis. These data thus confirmed that IL-13 > 15-LO>13(S)HpODE > PPARγ>MAO-A > ROS > p53>p21 axis has a major contribution in regulating cancer cell apoptosis and further identified 13(S)HpODE as a potential chemo-preventive agent which can improve the efficacy of cancer treatment as a combination compound.

Intermittent preventive treatment with Sulfadoxine pyrimethamine for malaria: a global overview and challenges affecting optimal drug uptake in pregnant women.

Malaria in Pregnancy (MiP) leading to morbidity and mortality is a major public health problem that poses significant risk to pregnant women and their fetus. To cope with this alarming situation, administration of Sulfadoxine-pyrimethamine (SP) drugs to pregnant women as an intermittent preventive treatment (IPT) from 16 weeks of gestation is recommended by the World Health Organization (WHO) guidelines. We conducted a comprehensive search of published articles related to MiP in last 10 years with predefined keywords or their synonyms. The mapping of malaria in pregnant women showed a prevalence rate up to 35% in many countries. Although IPTp-SP has been implemented in endemic regions since several years but the IPTp-SP coverage percentage vary from country to country and continue to remain below the target of 80%. Major reasons for low IPTp-SP involve gestational age at first prenatal visit, level of education, place of residence, knowledge of IPTp-SP benefits, and use of antenatal services. Several challenges including the emergence of septuple and octuple SP-resistant parasites is reported from many countries which make the prophylactic use of IPTp-SP currently debatable. This narrative review addresses the barriers for optimal use of IPTp-SP and discusses alternative approaches to increase the use and effectiveness of SP intervention for preventing MiP. The COVID pandemic has drastically affected the public health disrupting the management of diseases worldwide. In view of this, a brief summary of COVID impact on MiP situation is also included.

Molecular characterization of the severe falciparum malaria with typhoid co-infection: A case report.

Malaria and typhoid co-infections can be a serious public health issue in tropical countries leading to incorrect diagnosis due to overlapping clinical presentations of malaria and typhoid and hence, causing a delay in implementing the appropriate treatment regimen for these concurrent infections. This study reports a case of six-year-old female child co-infected with severe malaria (Plasmodium falciparum) and typhoid (Salmonella typhi) diagnosed by rapid malaria antigen test (RMAT) and blood culture respectively. Further, analysis of the chloroquine resistance gene Pfcrt for the falciparum demonstrated the presence of K76T mutant allele in pfcrt gene with high IC50 (150nM) for chloroquine (CQ) drug. The present case highlights the significance of timely identification and treatment of co-infections and also provides information about the circulating P. falciparum clinical strains.

Studying the disease severity in clinical isolates of Plasmodiumvivax.

Among the human malaria Plasmodium species, Plasmodium vivax is the most widespread species globally. In recent times, this historically benign species is now being recognized as also responsible for severe malaria infections in humans. Hence, a deeper insight of P.vivax immunopathogenesis in clinical patients is essential for malaria control and elimination strategies. Certain genes like vir genes, merozoite surface protein 3α genes (msp3α) and biomarkers like super oxide dismutase (SOD-1), tumor necrosis factor (TNF- α), interleukin (IL-10) are speculated to have some role in disease severity and thus can be useful as diagnostic markers. In the reported study, the clinical samples of P.vivax were genotyped for msp3α gene and cytokine analysis, expression profiling of vir genes were also carried out in these samples. A total of 84 P.vivax samples were collected (39 severe and 45 non-severe samples) and no correlation of parasitemia with severity of disease was seen in these samples (p-value = 0.38). On analysis four genotypes of msp3α were found, with type B (1.5 kb) as the predominant genotype. Cytokine analysis revealed SOD-1 and TNF-α levels to be significantly more in the severe group than in non-severe group, whereas for IL-10 no significant difference was observed between two clinical groups. The vir gene profiling revealed increased level of expression for vir-12, vir-14 related, and vir-17 like in severe group and vir-10 related gene expression was more in non-severe samples. There are multiple factors that bring phenotypic and genotypic changes in P.vivax malaria and thus, it is important to assess the potential diagnostic markers for detection of disease severity. In future, studies with more number of clinical samples should be undertaken for better insight of P.vivax disease severity.

Pancytopenia with hemophagocytic lymphohistiocytosis in Plasmodium falciparum: A unusual presentation.

Hematological manifestations such as anemia and thrombocytopenia are known complications in malaria. Here, we report two cases presented as pancytopenia with hepatosplenomegaly and initial diagnosis kept as hematological malignancy like leukemia but later on its diagnosed as malaria-associated hemophagocytic lymphohistiocytosis which is a rare entity. The aim of this report is to draw the attention of physicians, especially in tropical countries such as India and Sub-Saharan nations to keep in mind this uncommon presentation of malaria, though the exact pathophysiological mechanism still remains obscure.

3D printing of lignin: Challenges, opportunities and roads onward.

As the second most abundant biopolymer on earth, and as a resource recently becoming more available in separated and purified form on an industrial scale due to the development of new isolation technologies, lignin has a key role to play in transitioning our material industry towards sustainability. Additive manufacturing (AM), the most efficient-material processing technology to date, has likewise made great strides to promote sustainable industrial solutions to our needs in engineered products. Bringing lignin research to AM has prompted the emergence of the nascent "lignin 3D printing" field. This review presents the recent state of art of this promising field and highlights its challenges and opportunities. Following a review of the industrial availability, molecular attributes, and associated properties of technical lignins, we review R&D efforts at implementing lignin systems in extrusion-based and stereolithography (SLA) printing technologies. Doing so underlines the adage of lignin research that "all lignins are not created equal," and stresses the opportunity nested in this chemical diversity created mostly by differences in isolation conditions to molecularly select and tune the attributes of technical lignin systems towards desirable properties, be it by modification or polymer blending. Considering the AM design process in its entirety, we finally propose onward routes to bring the full potential to this emerging field. We hope that this review can help promote the unique value and overdue industrial role of lignin in sustainable engineered materials and products.

Epidemiology and clinical outcomes of severe Plasmodium vivax malaria in India.

OBJECTIVES: A systematic review and meta-analysis (SR-MA) of the available Indian literature on severe vivax malaria (SVM) was undertaken. METHODS: Relevant studies in eight electronic databases were retrieved and reviewed. The preferred reporting items for systematic reviews and meta-analyses (PRISMA) guidelines were followed. The methodological quality of the studies included in the MA was assessed. RESULTS: Overall, 162 studies were included in the work. The pooled proportion of SVM was 29.3%. The main severity signs/symptoms seen in SVM were jaundice, severe thrombocytopenia (ST), multi-organ dysfunction, and severe anaemia with pooled proportion of 37.4%, 37.2%, 24.2% and 20.4%, respectively. P. falciparum was inducing 6% less ST (RR = 0.94, 95% CI 0.5-1.5, I2 = 77.87%), 10% less thrombocytopenia (RR = 0.9, 95% CI 0.7-1.1, I2 = 91.68%) and 20% less DIC (RR = 0.8, 95% CI 0.3-1.9, I2 = 0%) than P. vivax. An atypical condition like myocarditis, was most commonly observed among the studied SVM cases. The mortality rate in SVM cases ranged from 0 to 12.9% among hospital patients with P. vivax mono-infections. CONCLUSIONS: The present SR-MA provides evidence for P. vivax as the etiologic agent of severe malaria leading to deaths in few cases as seen recently in India. However, research gaps outlined here emphasise the need for further studies on SVM in pregnancy, SVM in drug resistance and correlations with cytoadherence in disease severity due to P. vivax.

Artemisinin-based combination therapy (ACT) and drug resistance molecular markers: A systematic review of clinical studies from two malaria endemic regions - India and sub-Saharan Africa.

Artemisinin-based combination therapies (ACT) are currently used as a first-line malaria therapy in endemic countries worldwide. This systematic review aims at presenting the current scenario of drug resistance molecular markers, either selected or involved in treatment failures (TF) during in vivo ACT efficacy studies from sub-Saharan Africa (sSA) and India. Eight electronic databases were comprehensively used to search relevant articles and finally a total of 28 studies were included in the review, 21 from sSA and seven from India. On analysis, Artemether + lumefantrine (AL) and artesunate + sulfadoxine-pyrimethamine (AS + SP) are the main ACT in African and Indian regions with a 28-day efficacy range of 54.3-100% for AL and 63-100% for AS + SP respectively. It was observed that mutations in the Pfcrt (76T), Pfdhfr (51I, 59R, 108N), Pfdhps (437G) and Pfmdr1 (86Y, 184F, 1246Y) genes were involved in TF, which varied with respect to ACTs. Based on studies that have genotyped the Pfk13 gene, the reported TF cases, were mainly linked with mutations in genes associated with resistance to ACT partner drugs; indicating that the protection of the partner drug efficacy is crucial for maintaining the efficacy of ACT. This review reveals that ACT are largely efficacious in India and sSA despite the fact that some clinical efficacy and epidemiological studies have reported some validated mutations (i.e., 476I, 539T and 561H) in circulation in these two regions. Also, the role of PfATPase6 in ART resistance is controversial still, while P. falciparum plasmepsin 2 (Pfpm2) in piperaquine (PPQ) resistance and dihydroartemisinin (DHA) + PPQ failures is well documented in Southeast Asian countries but studied less in sSA. Hence, there is a need for continuous molecular surveillance of Pfk13 mutations for emergence of artemisinin (ART) resistance in these countries.

Switchgrass as oil and water-spill sorbent: Effect of particle size, torrefaction, and regeneration methods.

Switchgrass, both raw and torrefied, was tested for its ability to sorb water or oil. The cyclic performance was also examined, utilizing centrifugal extraction as the regeneration method. Both oil and water sorption capacity increase with the decreasing size of raw switchgrass particles. Results indicate that 3 mm raw switchgrass can sorb water at a capacity of about 6 times its mass and can sorb oil at a capacity of about 3 times its mass, which makes it a suitable biodegradable sorbent. Torrefaction at 220 °C for 30 min reduces water sorption capacity by an average of 55% but does not have a statistically significant impact on oil sorption. Sorption of liquid is negatively correlated to particle size. Centrifugation is able to partially desorb either liquid from the sorbent, and subsequent sorption cycles do not display lower sorption capacity than the first cycle when calculated on a dry mass basis.

Enhanced production of cordycepin in Ophiocordyceps sinensis using growth supplements under submerged conditions.

Cordycepin is a crucial bioactive compound produced by the fungus Cordyceps spp. Its therapeutic potential has been recognized for a wide range of biological properties such as anticancer, anti-diabetic, antidepressant, antioxidant, immunomodulation, etc. Moreover, its human random clinical trials depicted a promising anti-inflammatory activity that reduced the airway inflammation remarkably in asthmatic patients. But its overexploitation and low production of cordycepin in naturally growing biomass are insufficient to meet its existing market demand for its therapeutic use. Therefore, strategies for enhancement of cordycepin production in Cordyceps spp. are warranted. However, specifically, wild type Ophiocordyceps sinensis possesses a very low content of cordycepin and has restricted growth in natural mycelial biomass. To overcome these limitations, this study attempted to enhance cordycepin production in its mycelial biomass in vitro under submerged conditions by adding various growth supplements. The effect of these growth supplements was evaluated by reversed-phase high-performance liquid chromatography (RP-HPLC) which demonstrated that among nucleosides- hypoxanthine and adenosine; amino acids-glycine and glutamine; plant hormones- 1-naphthaleneacetic acid (NAA) and 3-indoleacetic acid (IAA); vitamin-thiamine (B1) from each group of growth supplements yielded a higher amount of cordycepin with 466.48 ±â€¯3.88, 380.23 ±â€¯1.78, 434.97 ±â€¯2.32, 269.78 ±â€¯2.92, 227.61 ±â€¯2.34, 226.02 ±â€¯1.69 and 185.26 ±â€¯2.35 mg/L respectively as compared to control with 13.66 ±â€¯0.64 mg/L. Further, at the transcriptional level, quantitative real time-polymerase chain reaction (qRT-PCR) analysis of genes associated with metabolism and cordycepin biosynthesis depicted significant upregulation of major downstream genes- NT5E, RNR, purA, and ADEK which corroborated well with RP-HPLC analysis. Taken together, the present study identified growth supplements as potential precursors to activate the cordycepin biosynthesis pathway leading to improved cordycepin production in O. sinensis.

Identification, Optimization of Culture Conditions, and Bioactive Potential of Chinese Caterpillar Mushroom Ophiocordyceps sinensis (Ascomycetes) Mycelium Isolated from Fruiting Body.

The present study deals with the challenges acquainted with in vitro culture of Ophiocordyceps sinensis. We have optimized the culture conditions for the growth of O. sinensis mycelium in semi-synthetic liquid media and determined antibacterial potential of the cultured mycelia extracts. In this study, mycelia were isolated from fruiting bodies and the isolate was identified as O. sinensis anamorph based on sequencing of internal transcribed spacer region. We investigated different culture conditions to optimize the growth of mycelia. Through this investigation, the isolated strain was observed to have its optimum growth at temperature (20°C), which yielded biomass of 12.38 g/L and pH (6.0) yielded biomass of 11.24g/L. Further to augment the production of mycelia, different carbon and nitrogen sources were optimized for mycelium growth in liquid media, out of which sucrose and corn steep powder proved to be the best carbon and nitrogen sources yielding biomass 14.01 g/L and 14.14 g/L, respectively. The evaluation of aqueous and methanolic extracts for antibacterial activity depicted that these extracts are active against all bacterial strains tested here. Aqueous extract depicted minimum inhibitory concentration (MIC) of 0.312, 0.019, 0.078, 0.312, and 0.625 mg/mL and methanolic extract depicted 1.25, 0.078, 0.009, 1.25, and 0.156 mg/mL against Pseudomonas aeruginosa, Escherichia coli, Bacillus cereus, Staphylococcus aureus, and Listeria monocytogenes, respectively. These results led to optimization of enhanced biomass production of O. sinensis, which can be a better alternative approach for further physiological studies and large-scale cultivation of this mushroom for its utilization for therapeutics and nutraceutical values.