Multi-locus sequence analysis reveals phylogenetically segregated Entamoeba histolytica population.

Amoebiasis, caused by the enteric parasite, Entamoeba histolytica, is one of the major food- and water-borne parasitic diseases in developing countries with improper sanitation and poor hygiene. Infection with E. histolytica has diverse disease outcomes, which are determined by the genetic diversity of the infecting strains. Comparative genetic analysis of infecting E. histolytica strains associated with differential disease outcomes from different geographical regions of the world is important to identify the specific genetic patterns of the pathogen that trigger certain disease outcomes of Amoebiasis. The strategy is able to elucidate the genealogical relation and population structure of infecting E. histolytica strains from different geographical regions. In the present study, we have performed a comparative genetic analysis of circulating E. histolytica strains identified from different parts of the world, including our study region, based on five tRNA-linked short tandem repeat (STR) loci (i.e., D-A, NK2, R-R, STGA-D and A-L) and evaluated their potential associations with differential disease outcomes of Amoebiasis. A number of regional-specific, emerging haplotypes of E. histolytica, significantly associated with specific disease outcomes have been identified. Haplotypes, which have a significant positive association with asymptomatic and amoebic liver abscess outcomes, showed a significant negative association with diarrheal outcome, or vice versa. Comparative multi-locus analysis revealed that E. histolytica isolates from our study region are phylogenetically segregated from the isolates of other geographical regions. This study provides a crucial overview of the population structure and emerging pattern of the enteric parasite, E. histolytica.

Short tandem repeat (STR) based sequence typing of Entamoeba histolytica identifies STGA-D locus as a genetic marker, associated with disease outcomes.

Amoebiasis, caused by the enteric parasite Entamoeba histolytica has differential disease outcomes. The association of parasite genotypes with outcomes of amoebic infection is still a paradox and requires to be explored. The genetic information of infecting strains from endemic settings of different geographical regions is essential to evaluate the relation. Comparative genetics of E. histolytica clinical isolates from different disease outcomes have been explored based on two tRNA-linked STR loci (STGA-D and A-L). All of the repeat patterns in the A-L locus were newly identified and unique to Indian isolates. The majority of newly identified repeat patterns in STGA-D locus have outcome-specific distributions, predicting the emergence of disease-specific mutations in this target locus. Statistical analysis further reinforces this observation, as identified repeat patterns only from STGA-D but not A-L locus were significantly associated with disease outcomes. Phylogenetic analysis indicates independent segregation and divergence of tRNA-linked STR arrays for each STR locus.

Non-vesicular Lipid Transport Machinery in Trichomonas vaginalis: Novel Drug Targets against Trichomoniasis.

Eukaryotic cells have separate membrane-enclosed organelles with distinct biochemical identities and specialized functions. The unique characteristics of each organelle are regulated by asymmetric distribution and intra-cellular trafficking of two important biomolecules, proteins and lipids. Non-vesicular lipid transport facilitated by lipid transfer proteins performs essential roles in intracellular lipid trafficking and homeostasis, while vesicular transport regulates protein trafficking. Comparative analysis of lipid transport machinery in protists could help us to understand the basis of parasitism and insight into eukaryotic evolution. Trichomonas vaginalis, a parasitic protist, greatly depends on receptor-ligand-mediated signaling pathways for cellular differentiation, nutrient uptake, secretion of virulence factors, and pathogenesis. Lipids, despite being key molecules of signaling cascades, have mechanisms of intracellular transport that are largely unexplored in T. vaginalis. We have identified a repertoire of seventeen potential lipid transfer protein (LTP) homologs in T. vaginalis based on a domain-based search on TrichDB (genome database of Trichomonas) coupled with bioinformatics analyses, which indicates the presence of well-organized lipid transport machinery in this parasite. We emphasized here their evolutionary uniqueness and conservation and discussed their potential implications for parasite biology in regard to future therapeutic targets against Trichomoniasis.

Persistence of behavioral abnormalities following corticosteroid therapy in children with initial episode of idiopathic nephrotic syndrome: a prospective longitudinal observation / Persistência de anormalidades comportamentais após corticoterapia em crianças com episódio inicial de síndrome nefrótica idiopática: uma observação longitudinal prospectiva

Abstract Introduction: Treatment of nephrotic syndrome with corticosteroid can cause several side- effects including behavioral abnormalities. The objectives of the study were to observe the proportion of non-relapsers having persistence of behavioral abnormalities after completion of treatment of initial episode and compare the abnormalities with relapsers, and to determine risk factors for persistence. Methods: Seventy-five children with a first episode of idiopathic nephrotic syndrome and 60 normal children were rated by parents for behavioral problems using the Child Behavior Checklist. The Parenting Stress Index was also evaluated. The children were rated before treatment and 12 and 36 weeks after. Results: Both relapsers and non-relapsers showed abnormalities in internalizing and externalizing domains at 12 weeks of steroid therapy. Non-relapsers had abnormal scores in the internalizing domain in 63.5 % and externalizing domain in 48.1% of cases at 36 weeks. Relapsers had abnormal scores in all the three behavior domains, but a significantly higher proportion of relapsers had abnormal scores regarding total behavior (65.2% vs 28.8%, p<0.01) and child domains (100% vs 57.7%, p<0.001) of Parenting Stress Index in comparison to non-relapsers at 36 weeks. Occurrence of relapse increased the risk (odds ratio 5.76, 95% CI 1.35-10.76, p< 0.001) for persistence of abnormal total behavior at 36 weeks follow-up. Conclusion: Persistence of abnormalities was observed not only in relapsers but also in non-relapsers. Relapse was found to be a significant risk factor for persistence of abnormal behaviors in these patients.

Resumo Introdução: O tratamento da síndrome nefrótica com corticosteroide pode causar vários efeitos colaterais, incluindo anormalidades comportamentais. Os objetivos do estudo foram observar a proporção de não-recidivos com persistência de anormalidades comportamentais após conclusão do tratamento do episódio inicial, comparar as anormalidades com os recidivos, e determinar fatores de risco para persistência. Métodos: 75 crianças com primeiro episódio de síndrome nefrótica idiopática e 60 crianças normais foram avaliadas pelos pais por problemas comportamentais usando o Checklist de Comportamento Infantil. O Índice de Estresse Parental também foi avaliado. As crianças foram avaliadas antes do tratamento, 12 e 36 semanas após. Resultados: Tanto recidivos quanto não recidivos mostraram anormalidades nos domínios de internalização e externalização às 12 semanas de terapia com esteroides. Não-recidivos apresentaram pontuações anormais no domínio de internalização em 63,5%, e no domínio de externalização, em 48,1% dos casos em 36 semanas. Recidivos tiveram pontuações anormais em todos os três domínios de comportamento, mas uma proporção significativamente maior de recidivos apresentou pontuações anormais em relação ao comportamento total (65,2% vs 28,8%, p<0,01) e domínios infantis (100% vs 57,7%, p<0,001) do Índice de Estresse Parental em comparação com não recidivos às 36 semanas. A ocorrência de recidiva aumentou o risco (odds ratio 5,76, 95% IC 1,35-10,76, p< 0,001) de persistência de comportamento total anormal em 36 semanas de acompanhamento. Conclusão: A persistência de anormalidades foi observada não apenas em recidivos, mas também em não recidivos. A recidiva foi um fator de risco significativo para a persistência de comportamentos anormais nesses pacientes.

Persistence of behavioral abnormalities following corticosteroid therapy in children with initial episode of idiopathic nephrotic syndrome: a prospective longitudinal observation.

INTRODUCTION: Treatment of nephrotic syndrome with corticosteroid can cause several side- effects including behavioral abnormalities. The objectives of the study were to observe the proportion of non-relapsers having persistence of behavioral abnormalities after completion of treatment of initial episode and compare the abnormalities with relapsers, and to determine risk factors for persistence. METHODS: Seventy-five children with a first episode of idiopathic nephrotic syndrome and 60 normal children were rated by parents for behavioral problems using the Child Behavior Checklist. The Parenting Stress Index was also evaluated. The children were rated before treatment and 12 and 36 weeks after. RESULTS: Both relapsers and non-relapsers showed abnormalities in internalizing and externalizing domains at 12 weeks of steroid therapy. Non-relapsers had abnormal scores in the internalizing domain in 63.5 % and externalizing domain in 48.1% of cases at 36 weeks. Relapsers had abnormal scores in all the three behavior domains, but a significantly higher proportion of relapsers had abnormal scores regarding total behavior (65.2% vs 28.8%, p<0.01) and child domains (100% vs 57.7%, p<0.001) of Parenting Stress Index in comparison to non-relapsers at 36 weeks. Occurrence of relapse increased the risk (odds ratio 5.76, 95% CI 1.35-10.76, p< 0.001) for persistence of abnormal total behavior at 36 weeks follow-up. CONCLUSION: Persistence of abnormalities was observed not only in relapsers but also in non-relapsers. Relapse was found to be a significant risk factor for persistence of abnormal behaviors in these patients.

Species richness and ß-diversity patterns of macrolichens along elevation gradients across the Himalayan Arc.

Understanding the species richness and ß-diversity patterns along elevation gradients can aid in formulating effective conservation strategies particularly in areas where local anthropogenic stresses and climate change are quite significant as in the Himalaya. Thus, we studied macrolichen richness and ß-diversity along elevational gradients at three sites, namely Kashmir (2200 to 3800 m a.m.s.l), Uttarakhand (2000-3700 m a.m.s.l) and Sikkim (1700 to 4000 m a.m.s.l) which cover much of the Indian Himalayan Arc. In all, 245 macrolichen species belonging to 77 genera and 26 families were collected from the three sites. Only 11 species, 20 genera and 11 families were common among the three transects. Despite the differences in species composition, the dominant functional groups in the three sites were the same: foliose, fruticose and corticolous forms. The hump-shaped elevation pattern in species richness was exhibited by most of the lichen groups, though an inverse hump-shaped pattern was also observed in certain cases. ß-diversity (ßsor) based on all pairs of comparisons along an elevation gradient varied from 0.48 to 0.58 in Kashmir, 0.03 to 0.63 in Uttarakhand and 0.46 to 0.77 in Sikkim. The contribution of turnover to ß-diversity was more than nestedness at all the three transects. Along elevation ß-diversity and its components of turnover and nestedness varied significantly with elevation. While species turnover increased significantly along the elevation in all the three transects, nestedness decreased significantly in Kashmir and Sikkim transects but increased significantly in the Uttarakhand transect. Except for the Kashmir Himalayan elevation transect, stepwise ß-diversity and its components of turnover and nestedness did not vary significantly with elevation. The present study, the first of its kind in the Himalayan region, clearly brings out that macrolichen species richness, ß-diversity, and its components of turnover and nestedness vary along the elevation gradients across the Himalayan Arc. It also highlights that contribution of turnover to ß-diversity is higher in comparison to nestedness at all the three transects. The variations in species richness and diversity along elevation gradients underpin the importance of considering elevational gradients in planning conservation strategies.

Therapeutic Potential of α- and ß-Pinene: A Miracle Gift of Nature.

α- and ß-pinene are well-known representatives of the monoterpenes group, and are found in many plants' essential oils. A wide range of pharmacological activities have been reported, including antibiotic resistance modulation, anticoagulant, antitumor, antimicrobial, antimalarial, antioxidant, anti-inflammatory, anti-Leishmania, and analgesic effects. This article aims to summarize the most prominent effects of α- and ß-pinene, namely their cytogenetic, gastroprotective, anxiolytic, cytoprotective, anticonvulsant, and neuroprotective effects, as well as their effects against H2O2-stimulated oxidative stress, pancreatitis, stress-stimulated hyperthermia, and pulpal pain. Finally, we will also discuss the bioavailability, administration, as well as their biological activity and clinical applications.

Antimicrobial activity of Bulbothrix setschwanensis (Zahlbr.) Hale lichen by cell wall disruption of Staphylococcus aureus and Cryptococcus neoformans.

In the present study, antimicrobial activity of a common Himalayan lichen viz. Bulbothrix setschwanensis (Zahlbr.) Hale extract in three common solvents (acetone, chloroform and methanol) was evaluated against six bacterial and seven fungal clinical strains. The acetone extract showed promising antimicrobial activity against S. aureus (1.56 mg/mL) and C. neoformans (6.25 mg/mL). Further, GC-MS analysis revealed 2,3-bis(2-methylpentanoyloxy)propyl 2-methylpentanoate and Ethyl 2-[(2R,3R,4aR,8aS)-3-hydroxy-2,3,4,4a,6,7,8,8a-octahydropyrano [3,2-b]pyran-2-yl]acetate as the predominant compounds. The combination of acetone extract with antibacterial drugs [kanamycin (KAN), rifampicin (RIF)] and antifungal drugs [amphotericin B (Amp B) and fluconazole (FLC)] showed lysis of S. aureus and C. neoformans at non-inhibitory concentration (FICI values were 0.31 for KAN, 0.18 for RIF, 0.37 for Amp B and 0.30 for FLC, respectively). Notably, the acetone extract confirmed cell wall damage of both S. aureus and C. neoformans cells and was clearly visualized under scanning electron microscopy (SEM), flow cytometry and confocal microscopy. Besides this, the three extracts also have less significant cytotoxic activity at MIC concentrations against mammalian cells (HEK-293 and HeLa). This study for the first time suggests that the chemical compounds present in the acetone extract of B. setschwanensis could be used against S. aureus and C. neoformans infections.

Behavioral Problems in Indian Children with Epilepsy.

OBJECTIVE: To assess prevalence of behavioral problems in children with epilepsy. METHODS: This was a cross-sectional study of children with epilepsy, and normal controls enrolled between July 2013 to June 2015. Child Behavior Checklist (CBCL) was used as a tool to assess the behavior based on parents reported observation. RESULTS: There were 60 children with epilepsy in 2-5 years and 80 in 6-14 years age groups, and 74 and 83 unaffected controls, respectively. Mean CBCL scores for most of the domains in children of both age groups were significantly higher than controls. Clinical range abnormalities were mainly detected in externalizing domain (23.3%) in 2-5 years, and in both internalizing (21.2%) and externalizing (45%) domains in children of 6-14 years. Younger age of onset, frequency of seizures and duration of disease had significant correlation with behavioral problems in both the age groups. Antiepileptic drug polytherapy was significantly associated with internalizing problems in older children. CONCLUSION: Age at onset, frequency of seizures and duration of disease were found to be significantly associated with occurrence of behavioral problems.

Behavioural abnormalities in children with new-onset nephrotic syndrome receiving corticosteroid therapy: results of a prospective longitudinal study.

BACKGROUND: Corticosteroid therapy can cause behavioural abnormalities in children with nephrotic syndrome. The objective of this study was to explore the timing of the appearance of abnormalities in their first episode. METHODS: Forty-five children with a first episode of idiopathic nephrotic syndrome (30 aged 2-5 and 15 aged 6-14 years) were assessed for behavioural problems using the Child Behaviour Checklist (CBCL) before, and after 6 and 12 weeks of oral steroid treatment. Sixty healthy children were included as controls. RESULTS: In both age groups, marked abnormalities of externalising behaviour were noticed, specifically in the domains of aggressive behaviour and attention problems. Clinical range or borderline externalising abnormalities were present in 73% of the younger children and 60% of the schoolchildren after 6 weeks of treatment. In the schoolchildren, abnormal internalising behaviour was also noted at 6 weeks, in 40% at borderline level and in 20% within the clinical range. Elevated scores were observed for the anxious/depressed and withdrawn/depressed domains. Most changes persisted at the 12-week observation. CONCLUSIONS: Children of both age groups showed significant attention problems and aggressive and abnormal externalising behaviour within 6 weeks of starting treatment. Parents should be informed and counselled about this potential adverse effect of steroid therapy.

Cognitive and behaviour dysfunction of children with neurocysticercosis: a cross-sectional study.

Eighty-three confirmed cases of neurocysticercosis diagnosed as per modified delBrutto criteria were enrolled in the study (Group-I) to observe cognitive and behavioural changes. Controls consisted of two groups: children with idiopathic generalized tonic-clonic seizure (Group-II) and normal children with non-specific cough (Group-III). Cases and controls were subjected to cognitive and behaviour assessment. There was significant difference in the intelligence quotient (IQ) of cases in domains of visual perception, immediate recall, analysis synthesis and reasoning, verbal ability, memory and spatial ability. In the age group of 6-18 years, cases had significantly more behaviour problems than control without seizure, in domains of anxious depressed, withdrawn depressed, somatic problems, social problems and rule-breaking behaviour. Neurocysticercosis causes decline in cognitive function and behaviours in older children, which should be recognized early for appropriate management and to avoid undue parental anxiety.

Clinical profile of somatoform disorders in children.

OBJECTIVE: To study the clinical profile, socio-demographic features, psychosocial stressors and outcome of somatoform disorders in children METHODS: Children up to the age of 18 year presenting with unexplained physical symptoms over a period of 6 months were evaluated. A detailed history and physical examination was carried out. Appropriate investigations were undertaken to exclude organic causes. Diagnosis was made according to DSM-IV-TR criteria. RESULTS: Forty-five children were diagnosed with somatoform disorders during the study period. The prevalence was 0.5% and 0.9% among outdoor and indoor patients, respectively. Conversion disorder (48.9%) was the commonest followed by other somatoform disorders (26.7%). Pseudoseizures and fainting attacks in conversion disorder and pain abdomen and general body pain in somatoform disorder were the commonest symptoms. Male to female ratio was 2.2:1. Urban children (25) were represented more than rural children (20). Stress factors were identified in 71.1% patients, which included fear of school or examinations. Thirty-three patients (73.3%) remained asymptomatic after counseling whereas, 8 patients (17.6%) had relapse requiring further counseling. Four patients (8.8%) showed no improvement and needed psychiatric evaluation. CONCLUSIONS: Early referral to mental health professional is required to avoid unnecessary investigations and delay in diagnosis of somatoform disorders in children.

Behavioural abnormalities in children with nephrotic syndrome.

BACKGROUND: Glucocorticoid therapy in children with nephrotic syndrome can lead to many adverse effects including behavioural problems. The present study was undertaken to assess the changes in individual behaviour among different sub-groups of patients with idiopathic nephrotic syndrome (INS) and also to find out the relationship, if any, between different behavioural problems with cumulative dose of steroid therapy. METHODS: This was a prospective hospital-based study. We assessed behavioural patterns in 131 children and adolescents with steroid-responsive INS aged 1.5-15 years. Fifty healthy children matched for age and gender were included to serve as controls. The Achenbach Child Behaviour Checklist was used to assess individual behaviour. Patients were sub-grouped according to age (1.5-5 and 6-15 years) and disease status (first attack before and after 12-week prednisolone, infrequent relapser, frequent relapser/steroid-dependent). RESULTS: All groups had significantly elevated mean behavioural abnormality scores for dimensions assessed in both age groups, except rule-breaking behaviour. Besides sleep problems, frequent relapsers/steroid-dependent patients exhibited maximum scores in comparison to first attack and infrequent relapsers in the 1.5- to 5-year age group. Total and individual behavioural scores showed close associations with cumulative prednisolone dose in both groups. CONCLUSIONS: It is evident that nephrotic syndrome patients should be given due consideration in clinical practice for behavioural abnormalities especially after steroid therapy.