HIV, violence, blame and shame:Pathways of risk to internalized HIV stigma among South African adolescents living with HIV: Pathways

Introduction: Internalized HIV-stigma is a key risk factor for negative outcomes amongst adolescents living with HIV (ALHIV), including non-adherence to anti-retroviral treatment, loss- to-follow-up and morbidity. This study tested a theoretical model of multi-level risk pathways to internalized HIV stigma among South African ALHIV. Methods: From 2013 – 2015, a survey using total population sampling of ALHIV who had ever initiated anti-retroviral treatment (ART) in 53 public health facilities in the Eastern Cape, South Africa was conducted. Community-tracing ensured inclusion of ALHIV who were defaulting from ART or lost to follow-up. 90.1% of eligible ALHIV were interviewed (n=1060, 55% female, mean age = 13.8, 21% living in rural locations). HIV stigma mechanisms (internalized, enacted, and anticipated), HIV-related disability, violence victimization (physical, emotional, sexual abuse, bullying victimization) were assessed using well-validated self-report measures. Structural equation modeling was used to test a theoretically-informed model of risk pathways from HIV-related disability to internalized HIV-stigma. The model controlled for age, gender and urban/rural address. Results: Prevalence of internalized HIV stigma was 26.5%. As hypothesized, significant associations between internalized stigma and anticipated stigma, as well as depression were obtained. Unexpectedly, HIV-related disability, victimization, and enacted stigma were not directly associated with internalized stigma. Instead significant pathways were identified via anticipated HIV stigma and depression. The model fitted the data well (RMSEA=.023; CFI=.94; TLI=.95; WRMR=1.070). Conclusions: These findings highlight the complicated nature of internalized HIV stigma. Whilst it is seemingly a psychological process, indirect pathways suggest multi-level mechanisms leading to internalized HIV stigma. Findings suggest that protection from violence within homes, communities and schools may interrupt risk pathways from HIV-related health problems to psychological distress and internalized HIV stigma. This highlights the potential for interventions that do not explicitly target adolescents living with HIV but are sensitive to their needs

Access to Community-Based HIV Services Among Transgender Women in Cambodia: Findings from a National Survey.

BACKGROUND: Globally, the prevalence of HIV among transgender women is more than 40 times higher than the prevalence in the general reproductive-age adults. They also face intersecting barriers to health, social, and legal services due to their hidden and stigmatized nature. Despite the particular needs, data regarding the access to services among transgender populations is scant globally. This study aims to identify characteristics of transgender women in Cambodia that may determine their accessibility to community-based HIV services. METHODS: In the National Biological and Behavioral Survey 2016, a structured questionnaire was used for face-to-face interviews with 1375 sexually active transgender women recruited from the capital city and 12 other provinces using the Respondent-Driven Sampling method. Weighted multivariate regression analysis was conducted to explore factors associated with access to community-based HIV services. RESULTS: The mean age of the participants was 25.8 years (SD = 7.1), and 45.0% reported having received at least one community-based HIV service in the past three months. Compared to participants who reported not having been reached by any community-based HIV programs, participants who reported having been reached by the programs were significantly more likely to reside in an urban setting (AOR = 1.41, 95% CI = 1.01-1.96), to have used gender-affirming hormones (AOR = 1.50, 95% CI = 1.17-1.92), to have been tested for HIV in the past six months (AOR = 7.42, 95% CI = 5.78-9.53), and to have been arrested by police or other authorities because of their transgender identity (AOR = 1.55, 95% CI = 1.03-2.33). Participants who reported having been reached by community-based HIV programs were significantly less likely to report being in a receptive role (AOR = 0.34, 95% CI = 0.15-0.82), to use condoms consistently with non-commercial male partners (AOR = 0.72, 95% CI = 0.55-0.94), and to perceive that their co-workers were not supportive regarding their transgender identity (AOR = 0.57, 95% CI = 0.44-0.98). CONCLUSIONS: Despite the extensive expansion of community-based HIV programs, less than half of transgender women in this national survey had access to the services. Innovative strategies and culturally sensitive interventions should be put in place to reach and respond to the needs of sub-groups of transgender women who are less likely to be reached by the existing traditional approaches

Late embryogenesis abundant proteins: Structural characterisation and interaction with α-synuclein

Ressurection plants are extraordinary because of their ability to withstand long periods without water, enter a state of anhydrobiosis, and fully recover upon water arrival. Ramonda serbica is a relic and endemic species that belong to a very small group of desiccation-tolerant plants in Europe. Underlying physiological, molecular and morphological mechanisms that enable these plants to survive harsh environmental conditions have been an appealing subject to many researchers. Most of the genes responsible for this amazing ability are present in other plants, and this path of research where those genes could be activated in crops is growing much more attention because of the imminent crisis regarding food supplies in the near future. Key components involved in the response to dehydration in R. serbica plants were analysed through a comprehensive transcriptomic, proteomic, metabolite and photosynthetic study. Late embryogenesis abundant proteins play a significant role in the complex defence processes involved in desiccation tolerance. Defining physicochemical characteristics and specific physiological functions of late embryogenesis abundant proteins – LEAPs may lead to their applicability in other areas of research

Two contrasting late embryogenesis abounded protein family groups of Ramonda serbica Panc.

Ramonda serbica Panc. is an ancient resurrection plant, that survives a long desiccation period and fully recovers metabolic functions upon watering. The main characteristic of desiccationtolerant plant species is their ability to accumulate protective late embryogenesis abounded protein (LEAPs). To propose their role in R. serbica desiccation tolerance we structurally analysed LEAPs in hydrated and desiccated leaves. According to transcriptomics, 318 LEAPs were identified and classified into seven family groups based on protein BLAST analysis and conserved motifs (Pfam). The largest LEAPs belonged to the LEA2 and LEA4 protein family groups. We employed online tools to analyse physicochemical characteristics (Expasy, ProtParam, BioPython, GRAVY calculator), disorder propensity, and characterization protein structures (FELLS, JPred, SOPMA, PsiPred, Phyre2, Espritz-DisProt, Espritz-X, Iupred, TMHMM, +Heliquest). The most abundant, atypical LEA2 group containing 127, mostly hydrophobic proteins, was divided into five subgroups. Members of this group were predicted to fold into globular domains, β-barrel at the C-terminus, followed by transmembrane hydrophobic-helices and disordered N-terminal regions. Results indicated the possible involvement in the protection of the chloroplastic membranes. The LEA4 group exhibited an exceptionally high tendency to form amphipathic α-helices and simultaneously had a high disorder propensity. This group is made of 96 proteins, classified into 3 subgroups. The high content of polar and charged amino acids (lysine, glutamate, and aspartate) is characteristic of this group. Motifs corresponding to the R. serbica LEA4 protein family group folded into A-type α-helices that contained positive, negative, and hydrophobic surfaces. Based on previous knowledge, the possible functions of the LEA2 and LEA4 groups are discussed with significant implications on cell preservation technology and the improvement of crop drought tolerance

Late embryogenesis abundant (LEA) proteins in Ramonda serbica Panc identification, classification and structural characterization

An ancient resurrection plant Ramonda serbica Panc. is able to survive a long desiccation period and reestablish metabolic activity upon watering. A hallmark of desiccation tolerance in the resurrection species is the accumulation of protective late embryogenesis abundant proteins (LEAPs). These intrinsically disordered proteins (IDPs) may stabilize the correct structure of proteins and membranes during cellular dehydration. The aim of our study was to assess LEA genes’ expression levels in hydrated (HL) and desiccated leaves (DL) and to identify, characterise, and estimate the potential role of R. serbica LEAPs in desiccation tolerance. In total, 318 LEAPs from HL and DL were identified and classified into the seven LEA protein family groups ranging from LEA1-LEA5, seed maturation proteins (SMPs), and dehydrins (DEH). Analysis of the physicochemical properties, motif architecture, secondary structure, homology, and phylogenetic relationships demonstrated that R. serbica LEAPs greatly differed among the LEA family groups. The most abundant LEA2 proteins (mostly downregulated upon desiccation) exhibited lower hydrophilicity and propensity to fold into organised globular domains. Oppositely, hydrophilic LEA4 proteins tended to form amphipathic, A-type, α-helices. Most of desiccation-upregulated LEA genes encoded highly disordered DEH1, LEA1, LEA4.2, and LEA4.3 proteins. While dehydrins might chelate metals and bind DNA under water deficit, other ID LEAPs (e.g. LEA1, LEA3, LEA4) might participate in forming intracellular proteinaceous condensates or adopt amphipathic α-helical conformation, enabling them to stabilise desiccation-sensitive proteins and membranes. Taken together, possible functions of LEAPs are discussed with significant implications on drought tolerance improvement of crops grown in arid areas

Efficient production of highly purified Late Embryogenesis Abundant (LEA) protein from Arabidopsis thaliana by recombinant DNA technology

Late embryogenesis abundant (LEA) proteins are induced in cellular dehydration, such as freezing, drought, or desiccation. They can be involved in antioxidative defense, ion sequestration, and structural stabilization of both membranes and enzymes during freezing or drying, while by forming intracellular proteinaceous condensates they increase structural integrity and intracellular viscosity of cells during desiccation 1. The genome of the model plant Arabidopsis thaliana contains 51 genes encoding LEA proteins2. The majority of these LEA proteins (35%) belongs to Pfam LEA_4 (PF02987) family. In silico analysis suggested that these proteins are highly hydrophilic proteins with significant intrinsically disordered protein (IDP) properties. In order to evaluate structural properties and possible functions of LEA_4 protein family under different water content, a representative AtLEA25 protein (At2g42560, 635 aa), naturally located in the cytoplasm of seeds3 was obtained in Escherichia coli by recombinant DNA technology. Although this technology has been traditionally used to over-express and purify various globular proteins, numerous reports have shown that the IDPs, due to their structural plasicity are naturally highly susceptible to proteolytic cleavage. To conduct structural and functional studies we developed a robust method to produce highly purified (>95% pure) AtLEA25 with no detectable amount of protein breakdown products.Abstract: Serbian Biochemical Society, 10th Conference, Kragujevac, Serbia, 24.09.202

Characterization of the late embryogenesis abundant (LEA) proteins family in hydrated and desiccated Ramonda serbica Panc. leaves

Endemic plant species, Ramonda serbica is a resurrection plant that can tolerate extreme dehydration (desiccation, loss of 95% of cellular water) even over months. The accumulation of late embryogenesis abundant proteins (LEAPs) is a crucial step in the mechanism of desiccation tolerance. The role of LEAPs is not completely resolved, but they are accepted as intrinsically disordered proteins (IDPs). Based on previously established de novo transcriptome database of R. serbica leaves we identify around 160 members of LEA gene family. Identified LEAPs were classified into six groups: LEA 1-5 and seed maturation proteins (SMPs) according to protein family (Pfam) database. Based on multiple sequence alignment, secondary structure prediction and 3D structure modeling, we conducted LEA protein structure analysis. We showed that more than 50% of identified LEAPs exhibited a high propensity to form α-helices. As predicted by several bioinformatic tools, more than 70% of identified LEAPs were found to be highly disordered. Thus, these proteins are predicted to be disordered in solution, but they acquire a secondary, predominantly α-helical structure during drying, in contrast to globular proteins, which most often causes the loss of structure upon dehydration. By using molecular dynamic simulations, we identified the most favorable conformations of representative LEAPs and we have studied conformational transitions driven by the water scarcity. Structural characterization of LEAPs is a key to understand their function and regulation of their intrinsic structural disorder-to-order transition during desiccation as a requirement for biological function, in order to promote development of new therapeutic strategies in neurodegenerative disorders, cell preservation technology and the improvement of crop drought tolerance.Serbian Biochemical Society, 10th Conference, Kragujevac, Serbia, 24.09.202

Long Covid stigma: estimating burden and validating scale in a UK-based sample

Background Stigma can be experienced as perceived or actual disqualification from social and institutional acceptance on the basis of one or more physical, behavioural or other attributes deemed to be undesirable. Long Covid is a predominantly multisystem condition that occurs in people with a history of SARSCoV2 infection, often resulting in functional disability. This study aimed to develop and validate a Long Covid Stigma Scale (LCSS); and to quantify the burden of Long Covid stigma. Methods Data from the follow-up of a co-produced community-based Long Covid online survey using convenience non-probability sampling was used. Thirteen questions on stigma were designed to develop the LCSS capturing three domains-enacted (overt experiences of discrimination), internalised (internalising negative associations with Long Covid and accepting them as self-applicable) and anticipated (expectation of bias/poor treatment by others) stigma. Confirmatory factor analysis tested whether LCSS consisted of the three hypothesised domains. Model fit was assessed and prevalence was calculated. Results 966 UK-based participants responded (888 for stigma questions), with mean age 48 years (SD: 10.7) and 85% female. Factor loadings for enacted stigma were 0.70-0.86, internalised 0.75-0.84, anticipated 0.58-0.87, and model fit was good. The prevalence of experiencing stigma at least 'sometimes' and 'often/always' was 95% and 76% respectively. Anticipated and internalised stigma were more frequently experienced than enacted stigma. Those who reported having a clinical diagnosis of Long Covid had higher stigma prevalence than those without. Conclusion This study establishes a scale to measure Long Covid stigma and highlights common experiences of stigma in people living with Long Covid

Prospective associations between bullying victimisation, internalised stigma, and mental health in South African adolescents living with HIV

Background: Adolescents living with HIV may be at elevated risk of psychological problems, which are correlated with negative health outcomes. In cross-sectional research with HIV-affected adolescents, bullying victimisation and internalised HIV stigma have been associated with poorer psychological health. We extended these findings and tested longitudinal associations between bullying victimisation, internalised stigma, and mental health among adolescents living with HIV. We also tested whether relationships between bullying victimisation and psychological symptoms were mediated by internalised stigma. Method: Adolescents living with HIV (n = 1060, 10–19 years, 55% female), who had ever initiated HIV treatment in 53 public health facilities in the Eastern Cape, South Africa, were interviewed and followed up 18 months later (n = 995, 94% retention). Participants completed well-validated measures of depression, anxiety, posttraumatic stress, bullying victimisation, and internalised stigma. Results: After adjusting for baseline mental health and sociodemographic characteristics, baseline internalised stigma prospectively predicted poorer outcomes on all psychological measures