Effect of Entamoeba histolytica infection on gut microbial diversity and composition in diarrheal patients from New Delhi.

During amoebiasis, colonization of the gut by Entamoeba histolytica can lead to alterations of the host microbiota. In this study, we have compared the gut microbiota of patients of amoebiasis with healthy controls using 16S rRNA gene variable regions, (V1-V3) and (V3-V5), of the bacterial genome. From this 16S rRNA gene amplicon data, one paired-end and two single-end datasets were selected and compared by the number of OTUs obtained, sequence count, and diversity analysis. Our results showed that the V1-V3-paired-end dataset gave the maximum number of OTUs in comparison to the two single-end datasets studied. The amoebiasis samples showed a significant drop in richness in the alpha diversity measurements and lower intra group similarity compared to the healthy controls. Bacteria of genus Prevotella, Sutterella, and Collinsella were more abundant in healthy controls whereas Escherichia, Klebsiella, and Ruminococcus were more abundant in the E. histolytica-positive patients. All the healthy controls harbored bacteria belonging to Faecalibacterium, Prevotella, Ruminococcus, Subdoligranulum, and Escherichia genera while all the E. histolytica-positive patient samples contained genus Enterobacter. The compositional changes in the gut microbiome observed in our study indicated a higher prevalence of pathogenic bacteria along with a depletion of beneficial bacteria in E. histolytica-infected individuals when compared with healthy controls. These results underline the interplay between E. histolytica and the human gut microbiome, giving important inputs for future studies and treatments.

Southeastern United States Predictors of COVID-19 in Nursing Homes.

This study's aim was to determine nursing home (NH) and county-level predictors of COVID-19 outbreaks in nursing homes (NHs) in the southeastern region of the United States across three time periods. NH-level data compiled from census data and from NH compare and NH COVID-19 infection datasets provided by the Center for Medicare and Medicaid Services cover 2951 NHs located in 836 counties in nine states. A generalized linear mixed-effect model with a random effect was applied to significant factors identified in the final stepwise regression. County-level COVID-19 estimates and NHs with more certified beds were predictors of COVID-19 outbreaks in NHs across all time periods. Predictors of NH cases varied across the time periods with fewer community and NH variables predicting COVID-19 in NH during the late period. Future research should investigate predictors of COVID-19 in NH in other regions of the US from the early periods through March 2021.

County-Level Social Determinants of Health and COVID-19 in Nursing Homes, United States, June 1, 2020-January 31, 2021.

OBJECTIVES: Nursing homes are a primary setting of COVID-19 transmission and death, but research has primarily focused only on factors within nursing homes. We investigated the relationship between US nursing home-associated COVID-19 infection rates and county-level and nursing home attributes. METHODS: We constructed panel data from the Centers for Medicare & Medicaid Services (CMS) minimum dataset, CMS nursing home data, 2010 US Census data, 5-year (2012-2016) American Community Survey estimates, and county COVID-19 infection rates. We analyzed COVID-19 data from June 1, 2020, through January 31, 2021, during 7 five-week periods. We used a maximum likelihood estimator, including an autoregressive term, to estimate effects and changes over time. We performed 3 model forms (basic, partial, and full) for analysis. RESULTS: Nursing homes with nursing (0.005) and staff (0.002) shortages had high COVID-19 infection rates, and locally owned (-0.007) or state-owned (-0.025) and nonprofit (-0.011) agencies had lower COVID-19 infection rates than privately owned agencies. County-level COVID-19 infection rates corresponded with COVID-19 infection rates in nursing homes. Racial and ethnic minority groups had high nursing home-associated COVID-19 infection rates early in the study. High median annual personal income (-0.002) at the county level correlated with lower nursing home-associated COVID-19 infection rates. CONCLUSIONS: Communities with low rates of nursing home infections had access to more resources (eg, financial resources, staffing) and likely had better mitigation efforts in place earlier in the pandemic than nursing homes that had access to few resources and poor mitigation efforts. Future research should address the social and structural determinants of health that are leaving racial and ethnic minority populations and institutions such as nursing homes vulnerable during times of crises.

Mapping community-level determinants of COVID-19 transmission in nursing homes: A multi-scale approach.

Deaths from the COVID-19 pandemic have disproportionately affected older adults and residents in nursing homes. Although emerging research has identified place-based risk factors for the general population, little research has been conducted for nursing home populations. This GIS-based spatial modeling study aimed to determine the association between nursing home-level metrics and county-level, place-based variables with COVID-19 confirmed cases in nursing homes across the United States. A cross-sectional research design linked data from Centers for Medicare & Medicaid Services, American Community Survey, the 2010 Census, and COVID-19 cases among the general population and nursing homes. Spatial cluster analysis identified specific regions with statistically higher COVID-19 cases and deaths among residents. Multivariate analysis identified risk factors at the nursing home level including, total count of fines, total staffing levels, and LPN staffing levels. County-level or place-based factors like per-capita income, average household size, population density, and minority composition were significant predictors of COVID-19 cases in the nursing home. These results provide a framework for examining further COVID-19 cases in nursing homes and highlight the need to include other community-level variables when considering risk of COVID-19 transmission and outbreaks in nursing homes.

A Comparative Study of Effect of 0.25% Levobupivacaine with Dexmedetomidine versus 0.25% Levobupivacaine in Ultrasound-Guided Supraclavicular Brachial Plexus Block.

Context: Dexmedetomidine, an α2-agonist, has been studied widely as an adjuvant to local anesthetics in regional anesthesia techniques to enhance the quality and duration of analgesia (DOA). It was hypothesized that addition of dexmedetomidine 0.5 ug.kg‒1 to levobupivacaine would prolong the DOA. Aims: We aimed to evaluate the efficacy of dexmedetomidine as an adjuvant to levobupivacaine in supraclavicular brachial plexus block with respect to onset and duration of sensory and motor blockade, and duration of analgesia. Settings and Design: This was a prospective randomized double-blind study carried out at a tertiary hospital attached to medical college. Subjects and Methods: Sixty American Society of Anesthesiologists PS Class I and II patients aged between 18 and 60 years of either sex, undergoing elective upper-limb surgery lasting more than 30 min, were included in the study. They were randomly divided into two groups of thirty each to receive ultrasound-guided supraclavicular brachial plexus block. Group L was given nerve block with 20 mL of 0.25% levobupivacaine and 1 mL saline, and Group D received 20 mL of 0.25% levobupivacaine with 0.5 ug.kg‒1 of dexmedetomidine (diluted to volume of 1 mL). Onset time and duration of sensory and motor blockade, time to first rescue analgesia, and hemodynamic parameters were recorded. Statistical Analysis Used: Chi-square test for qualitative variables and Student's unpaired "t" test for continuous variables were used for statistical analysis. Results: The onset of sensory and motor blockade was 6.51 ± 0.77 min and 10.71 ± 0.34 min in Group D and 9.9 ± 0.45 and 15.93 ± 1.92 min in Group L, respectively (P < 0.005). DOA was 9.53 ± 0.29 h in Group D and 3.89 ± 0.30 h in Group L (P < 0.001). The duration of sensory and motor block was 9.14 ± 0.19 h and 8.55 ± 0.31 h in Group D and 6.15 ± 3.02 and 5.61 ± 2.98 h in Group L, respectively (P < 0.005). No adverse effects were observed in either of the groups. Conclusions: Addition of 0.5 ug.kg‒1 of dexmedetomidine to 20 mL 0.25% levobupivacaine in ultrasound guided (USG)-guided supraclavicular brachial plexus block shortens the onset time of sensory and motor blockade and prolongs duration of sensory and motor block and DOA.

Phagocytosis of Gut Bacteria by Entamoeba histolytica.

The protist parasite Entamoeba histolytica causes amoebiasis, a major public health problem in developing countries. Only a small fraction of patients infected with the parasite display invasive disease involving colon or extra intestinal tissues such as liver. E. histolytica exists as two distinct forms, cysts, the infective form, and trophozoites, that are responsible for disease pathology. The latter multiply in the large intestine occasionally causing disease. The large intestine in humans is populated by a number of different bacterial communities and amoebic cells grow in their midst using some as food material. Several studies have shown relationship between bacteria and E. histolytica growth and virulence. However, an understanding of this relationship in human gut environment is not clear. We have investigated the possibility that there may be specific interaction of amoeba with different bacteria present in the gut environment by using a metagenomic pipe line. This was done by incubating bacteria isolated from human fecal material with E. histolytica and then identifying the bacterial population isolated from amoebic cells using a rRNA based metagenomic approach. Our results show that the parasite prefers a few bacterial species. One of these species is Lactobacillus ruminus which has never shown to be associated with E. histolytica.

Optimisation of in silico derived 2-aminobenzimidazole hits as unprecedented selective kappa opioid receptor agonists.

Kappa opioid receptor (KOR) is an important mediator of pain signaling and it is targeted for the treatment of various pains. Pharmacophore based mining of databases led to the identification of 2-aminobenzimidazole derivative as KOR agonists with selectivity over the other opioid receptors DOR and MOR. A short SAR exploration with the objective of identifying more polar and hence less brain penetrant agonists is described herewith. Modeling studies of the recently published structures of KOR, DOR and MOR are used to explain the receptor selectivity. The synthesis, biological evaluation and SAR of novel benzimidazole derivatives as KOR agonists are described. The in vivo proof of principle for anti-nociceptive effect with a lead compound from this series is exemplified.

Differential expression and immunolocalization of antioxidant enzymes in Entamoeba histolytica isolates during metronidazole stress.

Entamoeba histolytica infections are endemic in the Indian subcontinent. Five to eight percent of urban population residing under poor sanitary conditions suffers from Entamoeba infections. Metronidazole is the most widely prescribed drug used for amoebiasis. In order to understand the impact of metronidazole stress on the parasite, we evaluated the expression of two antioxidant enzymes, peroxiredoxin and FeSOD, in Entamoeba histolytica isolates during metronidazole stress. The results reveal that, under metronidazole stress, the mRNA expression levels of these enzymes did not undergo any significant change. Interestingly, immunolocalization studies with antibodies targeting peroxiredoxin indicate differential localization of the protein in the cell during metronidazole stress. In normal conditions, all the Entamoeba isolates exhibit presence of peroxiredoxin in the nucleus as well as in the membrane; however with metronidazole stress the protein localized mostly to the membrane. The change in the localization pattern was more pronounced when the cells were subjected to short term metronidazole stress compared to cells adapted to metronidazole. The protein localization to the cell membrane could be the stress response mechanism in these isolates. Colocalization pattern of peroxiredoxin with CaBp1, a cytosolic protein, revealed that the membrane and nuclear localization was specific to peroxiredoxin during metronidazole stress.

Genomic distribution of SINEs in Entamoeba histolytica strains: implication for genotyping.

BACKGROUND: The major clinical manifestations of Entamoeba histolytica infection include amebic colitis and liver abscess. However the majority of infections remain asymptomatic. Earlier reports have shown that some E. histolytica isolates are more virulent than others, suggesting that virulence may be linked to genotype. Here we have looked at the genomic distribution of the retrotransposable short interspersed nuclear elements EhSINE1 and EhSINE2. Due to their mobile nature, some EhSINE copies may occupy different genomic locations among isolates of E. histolytica possibly affecting adjacent gene expression; this variability in location can be exploited to differentiate strains. RESULTS: We have looked for EhSINE1- and EhSINE2-occupied loci in the genome sequence of Entamoeba histolytica HM-1:IMSS and searched for homologous loci in other strains to determine the insertion status of these elements. A total of 393 EhSINE1 and 119 EhSINE2 loci were analyzed in the available sequenced strains (Rahman, DS4-868, HM1:CA, KU48, KU50, KU27 and MS96-3382. Seventeen loci (13 EhSINE1 and 4 EhSINE2) were identified where a EhSINE1/EhSINE2 sequence was missing from the corresponding locus of other strains. Most of these loci were unoccupied in more than one strain. Some of the loci were analyzed experimentally for SINE occupancy using DNA from strain Rahman. These data helped to correctly assemble the nucleotide sequence at three loci in Rahman. SINE occupancy was also checked at these three loci in 7 other axenically cultivated E. histolytica strains and 16 clinical isolates. Each locus gave a single, specific amplicon with the primer sets used, making this a suitable method for strain typing. Based on presence/absence of SINE and amplification with locus-specific primers, the 23 strains could be divided into eleven genotypes. The results obtained by our method correlated with the data from other typing methods. We also report a bioinformatic analysis of EhSINE2 copies. CONCLUSIONS: Our results reveal several loci with extensive polymorphism of SINE occupancy among different strains of E. histolytica and prove the principle that the genomic distribution of SINEs is a valid method for typing of E. histolytica strains.

Tubastatin, a selective histone deacetylase 6 inhibitor shows anti-inflammatory and anti-rheumatic effects.

Epigenetic modifications represent a promising new approach to modulate cell functions as observed in autoimmune diseases. Emerging evidence suggests the utility of HDAC inhibitors in the treatment of chronic immune and inflammatory disorders. However, class and isoform selective inhibition of HDAC is currently favored as it limits the toxicity that has been observed with pan-HDAC inhibitors. HDAC6, a member of the HDAC family, whose major substrate is α-tubulin, is being increasingly implicated in the pathogenesis of inflammatory disorders. The present study was carried out to study the potential anti-inflammatory and anti-rheumatic effects of HDAC6 selective inhibitor Tubastatin. Tubastatin, a potent human HDAC6 inhibitor with an IC50 of 11 nM showed significant inhibition of TNF-α and IL-6 in LPS stimulated human THP-1 macrophages with an IC50 of 272 nM and 712 nM respectively. Additionally, Tubastatin inhibited nitric oxide (NO) secretion in murine Raw 264.7 macrophages dose dependently with an IC50 of 4.2 µM and induced α-tubulin hyperacetylation corresponding to HDAC6 inhibition in THP-1 cells without affecting the cell viability. Tubastatin showed significant inhibition of paw volume at 30 mg/kg i.p. in a Freund's complete adjuvant (FCA) induced animal model of inflammation. The disease modifying activity of Tubastatin was also evident in collagen induced arthritis DBA1 mouse model at 30 mg/kg i.p. The significant attenuation of clinical scores (~70%) by Tubastatin was confirmed histopathologically and was found comparable to dexamethasone (~90% inhibition of clinical scores). Tubastatin showed significant inhibition of IL-6 in paw tissues of arthritic mice. The present work has demonstrated anti-inflammatory and antirheumatic effects of a selective HDAC6 inhibitor Tubastatin.

Transfusion related acute lung injury in a neonate.

Transfusion related acute lung injury (TRALI) is a life threatening and potentially fatal complication of blood component transfusion, which largely remains under- diagnosed and under-reported, especially in neonates. The present case aims to emphasize that TRALI should be kept as a differential diagnosis in all groups of patients, including neonates, who develop acute respiratory distress and fresh lung infiltrations in the chest radiograph within 6 h of blood component transfusion in the absence of evidence of volume overload or cardiac dysfunction. Its recognition is important in view of the associated illness and death, for instituting correct management, and for eliminating implicated donors from donor panels.

Establishing minimal clinically important differences for scores on the pediatric evaluation of disability inventory for inpatient rehabilitation.

BACKGROUND AND PURPOSE: Standardized pediatric assessment tools such as the Pediatric Evaluation of Disability Inventory (PEDI) numerically quantify changes during rehabilitation through test scores, but they are unable to provide client-specific information regarding important changes in function. The purpose of this study was to identify the smallest change in PEDI scores during inpatient rehabilitation that was considered to be a minimal clinically important difference (MCID) by physical therapists and other clinicians. SUBJECTS AND METHODS: A retrospective review was done of the medical charts of 53 children and youth (1-19 years of age) discharged from an inpatient rehabilitation hospital. Fifteen clinicians (5 physical therapists, 6 occupational therapists, and 4 speech and language pathologists) who were masked to the PEDI scores provided ratings of the magnitude of functional changes during inpatient rehabilitation using a Likert scale and a visual analog scale (VAS). Ratings by clinicians were reduced to 4 categories, including the MCID, and compared with PEDI change scores. RESULTS: The MCIDs ranged from 6 to 15 points (X=11.5, 95% confidence interval= +/- 2.8) for all PEDI scales. Likert scale and VAS ratings were correlated (tau =.73-.80). DISCUSSION AND CONCLUSION: Across all scales, PEDI change scores on the order of about 11% (0-100 scale) appear to be meaningful to clinicians during a child's or adolescent's inpatient rehabilitation. These data can serve as a starting point for interpreting group and individual changes on the PEDI during physical therapy intervention in inpatient rehabilitation.