Antigenic repertoire of Plasmodium vivax transmission-blocking vaccine candidates from the Indian subcontinent

<p>Abstract</p> <p>Background</p> <p>Genetic polymorphism is an inevitable component of a multistage infectious organism, such as the malaria parasite. By means of genetic polymorphism, parasite opts particular polymorph and reveals survival advantage. <it>Pvs25 </it>and <it>pvs28 </it>are sexual stage antigen genes, expressed at the ookinete stage inside the mosquito gut, and considered as potential transmission-blocking vaccine candidates. This study presents sequence variations in two important transmission blocking antigen genes <it>pvs25 and pvs28 </it>in the field isolates of <it>P. vivax </it>from the Indian subcontinent.</p> <p>Methods</p> <p>One hundred microscopically diagnosed <it>P. vivax </it>isolates were collected from five geographical regions of India. <it>Pvs25 and pvs28 </it>genes were PCR amplified and sequenced to assess sequence variation among field isolates.</p> <p>Results</p> <p>A total of 26 amino acid substitutions were observed in Pvs25 (10) and Pvs28 (16) among field isolates of <it>P. vivax</it>. Tandem repeat polymorphism observed in <it>pvs28 </it>shows 3-6 tandem repeats in the field isolates. Seven and eight novel amino acid substitutions were observed in Pvs25 and Pvs28, respectively in Indian isolates. Comparison of amino acid substitutions suggests that majority of substitutions observed in global isolates were also present in Indian subcontinent. A single haplotype was observed to be major haplotype among isolates of Delhi, Nadiad, Chennai and Panna except in isolates of Kamrup. Further, population comparison analyses suggest that <it>P. vivax </it>isolates inhabiting in north-eastern region (Kamrup) were distantly related with the isolates from remaining parts of the country. Majority of the amino acid substitutions observed in Indian isolates were more identical to the substitutions reported from isolates of Thailand and Bangladesh.</p> <p>Conclusion</p> <p>Study uncovered many new amino acid substitutions as well as a predominance of single haplotype in Indian subcontinent except in north-eastern region of the country. The amino acid substitutions data generated in this study from different geographical regions of the Indian subcontinent could be helpful in designing a more effective anti-malarial transmission-blocking vaccine.</p

Molecular epidemiology of Plasmodium vivax anti-folate resistance in India

<p>Abstract</p> <p>Background</p> <p>Sulphadoxine and pyrimethamine are anti-folate drugs that show synergistic anti-malarial effect. Point mutations in <it>dihydrofolate reductase </it>(<it>dhfr</it>) and <it>dihydropteorate synthatase </it>(<it>dhps</it>) cause anti-folate drug resistance phenotype in human malaria parasites. This study presents pattern of point mutations in <it>dhfr/dhps </it>genes among Indian sub-continent.</p> <p>Methods</p> <p>Microscopically diagnosed one hundred <it>Plasmodium vivax </it>field isolates were collected from five widely separated geographical regions of India. <it>Dhfr </it>and <it>dhps </it>genes were PCR amplified and sequenced. Previously published mutations data were collected and analyzed using Chi square test to identify geographical cluster of mutant/wild type genotypes.</p> <p>Results</p> <p>Sequence analysis revealed single (S58R), double (S58R/S117N) and quadruple (F57L/S58R/T61M/S117T/) point mutations at <it>dhfr </it>and single (A383G) to double (A383G/A553G) mutations at <it>dhps </it>in <it>P. vivax </it>field isolates. In addition, three new mutations were also observed at <it>dhfr</it>. Both, <it>dhfr </it>and <it>dhps </it>genes revealed tandem repeat variations in field isolates. <it>Dhps </it>revealed very low mutation frequency (14.0%) compared to <it>dhfr </it>(50.70%). Comparative analysis revealed a progressive increase in frequency of quadruple mutant <it>dhfr </it>genotype (p < 0.001) within five years in north-eastern state (Kamrup, Assam). Frequency of <it>dhfr </it>genotypes revealed three distinct geographical clusters of wild (northern India), double mutant (southern India), and quadruple mutant (north-eastern and island regions of India) on the Indian sub-continent.</p> <p>Conclusion</p> <p>Study suggests that SP may be susceptible to <it>P. vivax </it>in India, except Andaman and north-eastern state. The distinction of geographical regions with sensitive and resistant parasite phenotypes would be highly useful for designing and administering national anti-malarial drug policy.</p

Larvicidal activity of neem oil (Azadirachta indica) formulation against mosquitoes

<p>Abstract</p> <p>Background</p> <p>Mosquitoes transmit serious human diseases, causing millions of deaths every year. Use of synthetic insecticides to control vector mosquitoes has caused physiological resistance and adverse environmental effects in addition to high operational cost. Insecticides of botanical origin have been reported as useful for control of mosquitoes. <it>Azadirachta indica </it>(Meliaceae) and its derived products have shown a variety of insecticidal properties. The present paper discusses the larvicidal activity of neem-based biopesticide for the control of mosquitoes.</p> <p>Methods</p> <p>Larvicidal efficacy of an emulsified concentrate of neem oil formulation (neem oil with polyoxyethylene ether, sorbitan dioleate and epichlorohydrin) developed by BMR & Company, Pune, India, was evaluated against late 3^rdand early 4^thinstar larvae of different genera of mosquitoes. The larvae were exposed to different concentrations (0.5–5.0 ppm) of the formulation along with untreated control. Larvicidal activity of the formulation was also evaluated in field against <it>Anopheles</it>, <it>Culex</it>, and <it>Aedes </it>mosquitoes. The formulation was diluted with equal volumes of water and applied @ 140 mg <it>a.i</it>./m²to different mosquito breeding sites with the help of pre calibrated knapsack sprayer. Larval density was determined at pre and post application of the formulation using a standard dipper.</p> <p>Results</p> <p>Median lethal concentration (LC₅₀) of the formulation against <it>Anopheles stephensi</it>, <it>Culex quinquefasciatus </it>and <it>Aedes aegypti </it>was found to be 1.6, 1.8 and 1.7 ppm respectively. LC₅₀values of the formulation stored at 26°C, 40°C and 45°C for 48 hours against <it>Ae. aegypti </it>were 1.7, 1.7, 1.8 ppm while LC₉₀values were 3.7, 3.7 and 3.8 ppm respectively. Further no significant difference in LC₅₀and LC₉₀values of the formulation was observed against <it>Ae. aegypti </it>during 18 months storage period at room temperature. An application of the formulation at the rate of 140 mg <it>a.i</it>./m²in different breeding sites under natural field conditions provided 98.1% reduction of <it>Anopheles </it>larvae on day 1; thereafter 100% reduction was recorded up to week 1 and more than 80% reduction up to week 3, while percent reduction against <it>Culex </it>larvae was 95.5% on day 1, and thereafter 80% reduction was achieved up to week 3. The formulation also showed 95.1% and, 99.7% reduction of <it>Aedes </it>larvae on day 1 and day 2 respectively; thereafter 100% larval control was observed up to day 7.</p> <p>Conclusion</p> <p>The neem oil formulation was found effective in controlling mosquito larvae in different breeding sites under natural field conditions. As neem trees are widely distributed in India, their formulations may prove to be an effective and eco-friendly larvicide, which could be used as an alternative for malaria control.</p

Anti-malarial activity of Holarrhena antidysenterica and Viola canescens, plants traditionally used against malaria in the Garhwal region of north-west Himalaya

<p>Abstract</p> <p>Background</p> <p>The increasing number of multidrug-resistant <it>Plasmodium </it>strains warrants exploration of new anti-malarials. Medicinal plant research has become more important, particularly after the development of Chinese anti-malarial drug artemisnin from <it>Artemisia annua</it>. The present study shows evaluation of anti-malarial effects of two plants commonly used against malaria in the Garhwal region of north-west Himalaya, in order to discover the herbal-based medicine.</p> <p>Methods</p> <p><it>In vitro </it>anti-plasmodial sensitivity of plant extracts was assessed using schizont maturation and parasite lactate dehydrogenase (pLDH) assay. Cytotoxic activities of the examined extracts were determined on L-6 cells of rat skeletal muscle myoblast. The 4-day test for anti-malarial activity against a chloroquine sensitive <it>Plasmodium berghei </it>NK65 strain in Swiss albino mice was used for monitoring <it>in vivo </it>activity of plant extracts.</p> <p>Results</p> <p>Chloroform extract of <it>H. antidysenterica </it>(HA-2) and petroleum ether extract of <it>V. canescens </it>(VC-1) plants significantly reduced parasitaemia in <it>P. berghei </it>infected mice. The extract HA-2 showed <it>in vitro </it>anti-plasmodial activity with its IC₅₀value 5.5 μg/ml using pLDH assay and ED₅₀value 18.29 mg/kg in <it>P. berghei </it>infected Swiss albino mice. Similarly petroleum ether extract of <it>V. canescens </it>(VC-1) showed <it>in vitro </it>anti-plasmodial activity with its IC₅₀value 2.76 μg/ml using pLDH assay and ED₅₀15.8 mg/kg in <it>P. berghei </it>infected mice. The extracts coded as HA-2 at 30 mg/kg and VC-1 at 20 mg/kg exhibited parasite inhibition in mice: 73.2% and 63.0% respectively. Of these two plant extracts, petroleum ether extract of <it>V. canescens </it>was found slightly cytotoxic.</p> <p>Conclusion</p> <p>The present investigation reflects the use of these traditional medicinal plants against malaria and these plants may work as potential source in the development of variety of herbal formulations for the treatment of malaria.</p

Global, regional, and national burden of disorders affecting the nervous system, 1990–2021: a systematic analysis for the Global Burden of Disease Study 2021

BackgroundDisorders affecting the nervous system are diverse and include neurodevelopmental disorders, late-life neurodegeneration, and newly emergent conditions, such as cognitive impairment following COVID-19. Previous publications from the Global Burden of Disease, Injuries, and Risk Factor Study estimated the burden of 15 neurological conditions in 2015 and 2016, but these analyses did not include neurodevelopmental disorders, as defined by the International Classification of Diseases (ICD)-11, or a subset of cases of congenital, neonatal, and infectious conditions that cause neurological damage. Here, we estimate nervous system health loss caused by 37 unique conditions and their associated risk factors globally, regionally, and nationally from 1990 to 2021.MethodsWe estimated mortality, prevalence, years lived with disability (YLDs), years of life lost (YLLs), and disability-adjusted life-years (DALYs), with corresponding 95% uncertainty intervals (UIs), by age and sex in 204 countries and territories, from 1990 to 2021. We included morbidity and deaths due to neurological conditions, for which health loss is directly due to damage to the CNS or peripheral nervous system. We also isolated neurological health loss from conditions for which nervous system morbidity is a consequence, but not the primary feature, including a subset of congenital conditions (ie, chromosomal anomalies and congenital birth defects), neonatal conditions (ie, jaundice, preterm birth, and sepsis), infectious diseases (ie, COVID-19, cystic echinococcosis, malaria, syphilis, and Zika virus disease), and diabetic neuropathy. By conducting a sequela-level analysis of the health outcomes for these conditions, only cases where nervous system damage occurred were included, and YLDs were recalculated to isolate the non-fatal burden directly attributable to nervous system health loss. A comorbidity correction was used to calculate total prevalence of all conditions that affect the nervous system combined.FindingsGlobally, the 37 conditions affecting the nervous system were collectively ranked as the leading group cause of DALYs in 2021 (443 million, 95% UI 378–521), affecting 3·40 billion (3·20–3·62) individuals (43·1%, 40·5–45·9 of the global population); global DALY counts attributed to these conditions increased by 18·2% (8·7–26·7) between 1990 and 2021. Age-standardised rates of deaths per 100 000 people attributed to these conditions decreased from 1990 to 2021 by 33·6% (27·6–38·8), and age-standardised rates of DALYs attributed to these conditions decreased by 27·0% (21·5–32·4). Age-standardised prevalence was almost stable, with a change of 1·5% (0·7–2·4). The ten conditions with the highest age-standardised DALYs in 2021 were stroke, neonatal encephalopathy, migraine, Alzheimer's disease and other dementias, diabetic neuropathy, meningitis, epilepsy, neurological complications due to preterm birth, autism spectrum disorder, and nervous system cancer.InterpretationAs the leading cause of overall disease burden in the world, with increasing global DALY counts, effective prevention, treatment, and rehabilitation strategies for disorders affecting the nervous system are needed

Sulphadoxine concentrations in plasma, red blood cells and whole blood in healthy and Plasmodium falciparum malaria cases after treatment with Fansidar using high-performance liquid chromatography

A reversed-phase high-performance liquid chromatographic method using acetonitrile-methanol-(1M) perchloric acid-water (30:9:0.8:95, v/v/v/v) at a flow of 1.5 ml min&#8722;1 on &#956;-Bondapak C18 column with UV (254 nm) detection has been developed for the separation of sulphadoxine, sulphalene and sulphamethoxazole from other antimalarials. Calibration curves were linear in the range 0.5-100 &#956;g ml&#8722;1. The limit of quantitation was 50 ng ml&#8722;1. Within-day and day-to-day coefficients of variation averaged 2.1 and 6.45%, respectively. The extraction recovery of sulphadoxine from plasma, red blood cells and whole blood was 90.28, 92.05 and 94.69%, respectively. Tle method has been used for the determination of sulphadoxine concentrations in plasma, red blood cells and whole blood of eight healthy and 50 Plasmodium falciparum malaria cases after administration of two tablets of Fansidar. Mean sulphadoxine concentration in plasma was higher than red blood cells or whole blood. Sulphadoxine concentration in plasma and whole blood of P. falciparum malaria cases was significantly higher as compared to healthy volunteers while it was the same in red blood cells. Sulphadoxine was absorbed much less in red blood cells than in plasma or whole blood

Determination of sulfalene in plasma, red blood cells and whole blood by high-performance liquid chromatography

A normal phase high-performance liquid chromatographic method using dichloromethane-methanol-perchloric acid (1 M) (96:9:1, v/v) at a flow-rate of 1 ml/min on a Nucleosil 100<SUP>−7</SUP> column (250 × 8 × 4 mm) and UV detection at 254 nm, has been developed to determine the concentration of sulfalene in plasma, red blood cells and whole blood after oral administration of the antimalarial drug metakelfin. The coefficient of variation was 7.1% and the extraction recovery was 82%. Mean concentrations of sulfalene on days 1, 7 and 15 were: 49.56, 10.46 and 2.24 μg/ml in plasma, 25.02, 4.34 and 0.84 μg/ml in red blood cells and 21.12, 4.44 and 1.00 μg/ml in whole blood, respectively. Quinine, chloroquine, desethylchloroquine, mefloquine, primaquine, sulfadoxine, pyrimethamine and dapsone did not interfere in the detection of sulfalene