In situ characterization of immune cells mediating potential control of HIV infection in the female genital mucosa

Heterosexual HIV transmission is the most common viral transmission route, worldwide, where young women are more susceptible to HIV infection than men. To establish a persistent infection the virus needs to cross the mucosal surface of the female genital tract (FGT). The FGT mucosa is thus considered to be the portal of HIV entry and initial site of viral replication. A better understanding of the immunological milieu at the portal of viral entry is crucial for the development of preventive interventions. Hence, in this thesis we quantified and characterized immune cells, primarily subsets of CD8+ T cell, in the FGT tissues of HIV-infected and uninfected women. In Paper I and II, we observed that HIV-infected women displayed similar levels of CD4 expression in their ectocervix as compared to uninfected controls. However, they showed higher immune activation levels as well as signs of HIV replication in their cervix. An inflammatory environment in the genital mucosa may promote viral replication and genital shedding and thereby increase the risk of sexual HIV transmission. The HIV-infected women also had elevated levels of cervical CD8+ cells, which correlated significantly with cervical viral load. The CD8+ cells were predominantly intraepithelial CD8+ T cells and 60% of them expressed CD103 (Paper II, III). Our data thus suggests that CD8+ T cells, including tissue-resident CD8+ T cells, may be actively recruited to/or expanded in the genital mucosa of chronically HIV-infected sex workers. This implies that tissue-residing CD8+ T cells play an important role in local HIV pathogenesis. To further investigate immune responses in the FGT mucosa we assessed the expression of MAIT cells in the genital tissues of healthy women (Paper IV). FGT-derived MAIT cells preferentially localized in the ectocervical epithelium and were biased towards IL-17 and IL-22 production upon bacterial stimulation. This indicates that functional MAIT cells localized near the luminal surface of the genital mucosa may be important for the preservation of the genital barrier integrity and may act as a first line of defence against invading pathogens. In summary, this thesis describes the localization and distribution of immune cells in the genital mucosa of HIV-infected and uninfected women. Studies described here may contribute to the knowledge needed for development of vaccination and/or microbicide strategies against HIV and other sexually transmitted infections

Control for interfaces and handling qualities for space telerobotic vehicles

Thesis (M.S.)--Massachusetts Institute of Technology, Dept. of Aeronautics and Astronautics, 1991.Includes bibliographical references (leaf 63).by Anna Gibbs Cinniger.M.S

How do parents frame their engagement experience in early intervention? A grounded theory study

PURPOSE: Parent and therapist engagement and partnership are critical in early intervention physiotherapy and occupational therapy for infants with cerebral palsy to improve outcomes. The main aim of this study was to understand how parents perceive their engagement experience in early intervention over time. METHODS: Grounded theory methodology was used. Twenty parents of diverse backgrounds participated in 22 interviews (including some repeated longitudinally) to reflect on their engagement experience within the context of early intervention community services provided in the UK NHS. RESULTS: The findings highlight how parents' perspectives of their engagement in EI change according to critical circumstances, including their preceding neonatal trauma, the at-risk CP label, firmer diagnosis of CP and their child's response to intervention. We theorise that this disrupted transition experience to parenthood becomes part of parental framing (or sense-making) of their engagement in EI. Overlapping frames of uncertainty, pursuit and transformation capture and explain nuances in parents' engagement patterns within EI over time. CONCLUSION: This theorising has implications for early intervention therapists in how they engage in the lives of families and partner with parents to support healthier parental transition, wellbeing and subsequent improved infant outcomes

Methods for the evaluation of the Jamie Oliver Ministry of Food program, Australia.

BACKGROUND: Community-based programs aimed at improving cooking skills, cooking confidence and individual eating behaviours have grown in number over the past two decades. Whilst some evidence exists to support their effectiveness, only small behavioural changes have been reported and limitations in study design may have impacted on results.This paper describes the first evaluation of the Jamie Oliver Ministry of Food Program (JMoF) Australia, in Ipswich, Queensland. JMoF Australia is a community-based cooking skills program open to the general public consisting of 1.5 hour classes weekly over a 10 week period, based on the program of the same name originating in the United Kingdom. METHODS/DESIGN: A mixed methods study design is proposed. Given the programmatic implementation of JMoF in Ipswich, the quantitative study is a non-randomised, pre-post design comparing participants undergoing the program with a wait-list control group. There will be two primary outcome measures: (i) change in cooking confidence (self-efficacy) and (ii) change in self-reported mean vegetable intake (serves per day). Secondary outcome measures will include change in individual cooking and eating behaviours and psycho-social measures such as social connectedness and self-esteem. Repeated measures will be collected at baseline, program completion (10 weeks) and 6 months follow up from program completion. A sample of 250 participants per group will be recruited for the evaluation to detect a mean change of 0.5 serves a day of vegetables at 80% power (0.5% significance level). Data analysis will assess the magnitude of change of these variables both within and between groups and use sub group analysis to explore the relationships between socio-demographic characteristics and outcomes.The qualitative study will be a longitudinal design consisting of semi-structured interviews with approximately 10-15 participants conducted at successive time points. An inductive thematic analysis will be conducted to explore social, attitudinal and behavioural changes experienced by program participants. DISCUSSION: This evaluation will contribute to the evidence of whether cooking programs work in terms of improving health and wellbeing and the underlying mechanisms which may lead to positive behaviour change. TRIAL REGISTRATION: Australian and New Zealand Trial registration number: ACTRN12611001209987

Physiological synchrony and shared flow state in Javanese gamelan: positively associated while improvising, but not for traditional performance

The experience of shared flow refers to the optimal balance between challenge and ability for a given task, resulting from interpersonal action in a group situation. The performance of Javanese gamelan is an ideal setting to investigate shared flow, due to the requirement that all performers on varying instrumental parts work harmoniously, allowing for shared flow and its native equivalent, ngeli. To minimise the disruption of flow, while still measuring it continuously, one way to assess a person’s state is by measuring physiological responses of the sympathetic (i.e., fight-or-flight) system, namely heart rate and skin conductance. Flow has been related to physiological signatures, and shared actions in music-making have been related to synchronised physiology. However, to our knowledge, no study yet has directly investigated the links between shared physiology and shared flow. Therefore, this study aimed to assess the associations between flow states, physiological synchrony, and Javanese gamelan playing. Subsequently, we tested for differences between advanced and beginner groups playing traditional gamelan pieces and improvising. Firstly, a factor analysis revealed a two-factor solution of Awareness and Absorption for self-reported shared flow. Next, using inter-subject correlation to assess synchrony and circular shuffling to infer significance, we found a greater proportion of significance in traditional playing compared to improvised playing for the experienced group, and the opposite for the beginner group. Lastly, linear mixed models revealed largely positive associations between synchronised physiology and shared flow during improvised playing, and negative associations during traditional playing, regardless of experience levels. This study demonstrates methodological possibilities for the quantitative study of shared flow in music-making contexts, and potential differences in shared flow experience in improvised and traditional, or prescribed, playing

PIPITS: an automated pipeline for analyses of fungal internal transcribed spacer sequences from the Illumina sequencing platform

1. Studying fungal biodiversit y using data generated from Illumina MiSeq sequencing platforms poses a number of bioinformatic challenges with the analysis typically involving a large number of tools for each analytical step from quality filtering to generating identified operational taxonomic unit (OTU) abundance tables. 2. Here, we introduce PIPITS, an o pen-source stand-alone suite of software for automated processing of Illumina MiSeq sequences for fungal community analysis. PIPITS e xploits a number of state of the art applications to process paired-end reads from quality filtering to producing OTU abundance tables. 3. We pro vide detailed descriptions of the pipeline and show its utility in the analysis of 9 396 092 sequences generated on the MiSeq platform from Illumina MiSeq. 4. PIPITS is the first automated bioinformatics pipeline dedicated for fungal ITS sequences which incorporates ITSx to extract subregions of ITS and exploits the latest RDP Classifier to classify sequences against the curatedUNITE fungal data set

Transcriptional Regulation of Lipophorin Receptors Supports Neuronal Adaptation to Chronic Elevations of Activity

Activity-dependent modifications strongly influence neural development. However, molecular programs underlying their context and circuit-specific effects are not well understood. To study global transcriptional changes associated with chronic elevation of synaptic activity, we performed cell-type-specific transcriptome profiling of Drosophila ventral lateral neurons (LNvs) in the developing visual circuit and identified activity-modified transcripts that are enriched in neuron morphogenesis, circadian regulation, and lipid metabolism and trafficking. Using bioinformatics and genetic analyses, we validated activity-induced isoform-specific upregulation of Drosophila lipophorin receptors LpR1 and LpR2, the homologs of mammalian low-density lipoprotein receptor (LDLR) family proteins. Furthermore, our morphological and physiological studies uncovered critical functions of neuronal lipophorin receptors (LpRs) in maintaining the structural and functional integrities in neurons challenged by chronic elevations of activity. Together, our findings identify LpRs as molecular targets for activity-dependent transcriptional regulation and reveal the functional significance of cell-type-specific regulation of neuronal lipid uptake in experience-dependent plasticity and adaptive responses

Presence of CD8+ T Cells in the Ectocervical Mucosa Correlates with Genital Viral Shedding in HIV-Infected Women despite a Low Prevalence of HIV RNA–Expressing Cells in the Tissue

The female genital tract is a portal of entry for sexual HIV transmission and a possible viral reservoir. In this study, the ectocervical CD8+ T cell distribution was explored in situ and was related to expression of CD3 and HLA-DR and presence of HIV RNA. For this purpose, ectocervical tissue samples and genital secretions were collected from HIV-seropositive (HIV+) Kenyan female sex workers (FSWs) (n = 20), HIV-seronegative (HIV−) FSWs (n = 17), and HIV− lower-risk women (n = 21). Cell markers were assessed by in situ staining and by quantitative PCR. HIV RNA expression in tissue was analyzed by in situ hybridization, and viral shedding was assessed by quantitative PCR. The HIV+FSW group had a higher amount of total cells and CD8+, CD3+, and HLA-DR+ cells compared with the HIV−FSW group and HIV− lower-risk women. The majority of CD8+ cells were CD3+ T cells, and the numbers of CD8+ cells correlated significantly with plasma and cervical viral load. HIV RNA expression in situ was found in 4 of the 20 HIV+FSW women but did not correlate with cervical or plasma viral load. Thus, the HIV+ women displayed high numbers of CD8+, CD3+, and HLA-DR+ cells, as well as a limited number of HIV RNA+ cells, in their ectocervical mucosa; hence, this localization cannot be neglected as a potential viral reservoir. The elevated levels of CD8+ T cells may play a role in the immunopathogenesis of HIV in the female genital tract

Presence of CD8+ T Cells in the Ectocervical Mucosa Correlates with Genital Viral Shedding in HIV-Infected Women despite a Low Prevalence of HIV RNA–Expressing Cells in the Tissue

The female genital tract is a portal of entry for sexual HIV transmission and a possible viral reservoir. In this study, the ectocervical CD8+ T cell distribution was explored in situ and was related to expression of CD3 and HLA-DR and presence of HIV RNA. For this purpose, ectocervical tissue samples and genital secretions were collected from HIV-seropositive (HIV+) Kenyan female sex workers (FSWs) (n = 20), HIV-seronegative (HIV−) FSWs (n = 17), and HIV− lower-risk women (n = 21). Cell markers were assessed by in situ staining and by quantitative PCR. HIV RNA expression in tissue was analyzed by in situ hybridization, and viral shedding was assessed by quantitative PCR. The HIV+FSW group had a higher amount of total cells and CD8+, CD3+, and HLA-DR+ cells compared with the HIV−FSW group and HIV− lower-risk women. The majority of CD8+ cells were CD3+ T cells, and the numbers of CD8+ cells correlated significantly with plasma and cervical viral load. HIV RNA expression in situ was found in 4 of the 20 HIV+FSW women but did not correlate with cervical or plasma viral load. Thus, the HIV+ women displayed high numbers of CD8+, CD3+, and HLA-DR+ cells, as well as a limited number of HIV RNA+ cells, in their ectocervical mucosa; hence, this localization cannot be neglected as a potential viral reservoir. The elevated levels of CD8+ T cells may play a role in the immunopathogenesis of HIV in the female genital tract