Single-cell epigenomic dysregulation of Systemic Sclerosis fibroblasts via CREB1/EGR1 axis in self-assembled human skin equivalents.

Systemic sclerosis (SSc) is an autoimmune disease characterized by skin fibrosis, internal organ involvement and vascular dropout. We previously developed and phenotypically characterized an in vitro 3D skin-like tissue model of SSc, and now analyze the transcriptomic (scRNA-seq) and epigenetic (scATAC-seq) characteristics of this model at single-cell resolution. SSc 3D skin-like tissues were fabricated using autologous fibroblasts, macrophages, and plasma from SSc patients or healthy control (HC) donors. SSc tissues displayed increased dermal thickness and contractility, as well as increased α-SMA staining. Single-cell transcriptomic and epigenomic analyses identified keratinocytes, macrophages, and five populations of fibroblasts (labeled FB1 - 5). Notably, FB1 APOE-expressing fibroblasts were 12-fold enriched in SSc tissues and were characterized by high EGR1 motif accessibility. Pseudotime analysis suggests that FB1 fibroblasts differentiate from a TGF-ß1-responsive fibroblast population and ligand-receptor analysis indicates that the FB1 fibroblasts are active in macrophage crosstalk via soluble ligands including FGF2 and APP. These findings provide characterization of the 3D skin-like model at single cell resolution and establish that it recapitulates subsets of fibroblasts and macrophage phenotypes observed in skin biopsies.

Adnexal arteriovenous malformation presenting with spontaneous massive haemoperitoneum.

Arteriovenous malformations (AVMs) are vascular anomalies composed of a tangle of abnormal vessels in which one or more feeding arteries are directly connected to one or more draining veins via a nidus with no intervening capillary bed. The adnexa are particularly rare sites for the formation of such malformations. Here, we present the case of a middle-aged woman who presented with spontaneous massive haemoperitoneum occurring as a result of a ruptured adnexal AVM. The diagnosis was suspected on transabdominal sonography and confirmed on CT angiography. The patient was shifted to the interventional radiology suite for an urgent angioembolisation following which she improved haemodynamically and her symptoms resolved. The case highlights the fact that although exceedingly rare, gonadal AVMs are an important cause of spontaneous intraperitoneal bleeding. Diagnostic and interventional radiology play an important role in the early and accurate diagnosis of this entity, and angioembolisation can be lifesaving in such patients.

Prospective quantitative study: Doppler ultrasound in the evaluation of chronic renal allograft disease and correlation with histopathological finding.

Background and Objectives: Renal allograft biopsy is the gold standard for diagnosing chronic allograft nephropathy, but noninvasive methods are needed to avoid unnecessary biopsies. Doppler ultrasonography, particularly the resistive index (RI), correlates with renal allograft dysfunction. This study aims to assess the relationship between renal sonographic parameters and biochemical parameters in diagnosing graft interstitial fibrosis. Methods: The study evaluated 60 renal allograft recipients for sonographic renal morphological features and Doppler indices. The estimated glomerular filtration rate (eGFR) was calculated, and cortical fibrosis after the biopsy was determined using the Banff score. Continuous variables like mean and SD were calculated, and categorical variables were reported using frequencies and proportions. Associations were examined using independent sample t-tests, χ 2tests, and multivariate regression analysis. Results: The mean eGFR was 75.23±25.45 ml/min/1.73 m2. A significant correlation of eGFR with RI (r=0.341, P=0.008) was seen. A significant difference in mean RI (F=10.167; df=2,57; P<0.001) was seen among the histological grades of fibrosis. Among the histological grades of fibrosis, significant differences in RI among mild and moderate (S.E. 0.033, P=0.043), mild and severe (S.E. 0.026, P=0.001) as well as moderate and severe (S.E. 0.036, P=0.029) was seen. Conclusion: Doppler was able to noninvasively predict allograft fibrosis and could be used as a complementary imaging tool during the follow-up of renal allograft patients. Future research is needed to improve evidence, diagnostic criteria, guidelines, and long-term impact.

CDDO-Methyl Ester Inhibits BRAF Inhibitor Resistance and Remodels the Myeloid Compartment in BRAF-mutant Melanoma.

Approximately 50% of advanced melanomas harbor activating BRAF V600E mutations that are sensitive to BRAF inhibition. However, the duration of the response to BRAF inhibitors (BRAFi) has been limited due to the development of acquired resistance, which is preceded by recruitment of immunosuppressive myeloid cells and regulatory T cells (T regs ). While the addition of MAPK/ERK kinase 1 inhibitors (MEKi) prolongs therapeutic response to BRAF inhibition, most patients still develop resistance. Using a Braf V600E/+ /Pten -/- graft mouse model of melanoma, we now show that the addition of the methyl ester of the synthetic triterpenoid 2-cyano-3,12-dioxooleana-1,9(11)-dien-28-oic acid (C-Me) to the BRAFi vemurafenib analog PLX4720 at resistance significantly reduces tumor burden. Dual treatment remodels the BRAFi resistant-tumor microenvironment (TME), reducing infiltration of T regs and tumor associated macrophages (TAMs), and attenuates immunosuppressive cytokine production. For the first time, we characterize myeloid populations using scRNA-seq in BRAFi-resistant tumors and demonstrate that restoration of therapeutic response is associated with significant changes in immune-activated myeloid subset representation. Collectively, these studies suggest that C-Me inhibits acquired resistance to BRAFi. Use of C-Me in combination with other therapies may both inhibit melanoma growth and enhance therapeutic responsiveness more broadly.

Effect of Healthy Transitions intervention in improving family planning uptake among adolescents and young women in Western Nepal: A pre-and post-intervention study.

BACKGROUND: Contraceptive use can prevent unintended pregnancies, early childbearing, and abortion-related deaths. Despite these benefits, the use of modern contraceptives remains low among adolescent girls and young women (AGYW) in Nepal. To address this gap, the Healthy Transitions Project was implemented in Karnali Province, Nepal from February 2019 to September 2021. This study aimed at measuring the effect of Healthy Transitions' intervention on improving knowledge and use of modern family planning methods among AGYW in Nepal. METHODS: We used a pre- and post-intervention study design to assess the effect of Healthy Transitions project. A quantitative survey was conducted at baseline and after the first cohort of AGYW had completed the intervention (1 year later). The baseline survey was conducted in 2019 with a cohort of 786 married and unmarried AGYW aged 15-24 years. An end line survey was conducted in 2020 with 565 AGYW who were interviewed at baseline. Data analysis was done using STATA version 15.1. The exact McNemar significance probability value was used to decide the significance of difference between baseline and endline. RESULTS: The knowledge and uptake of modern family planning methods have increased in the endline compared to the baseline. AGYW recognised 10 out of the ten modern methods at endline, a significant increase from 7 at baseline (p<0.001). Among AGYW, 99% were aware of sources to obtain family planning methods, compared with 92% at baseline (p< 0.001). The proportion of married AGYW using modern contraceptive methods was significantly higher at the endline 33%, than baseline (26%) (p<0.001). CONCLUSION: Our results show that multilevel demand and supply-side interventions, targeting adolescents and young women, their families, community, and health system helped to improve knowledge and use of modern family planning methods among AGYW. The study suggests that these intervention approaches can be adopted to improve family planning use among adolescents and young women in other similar settings.

Findings from a mixed-methods evaluation of a multi-level adolescent and youth reproductive and maternal health intervention in Karnali Province, Nepal.

INTRODUCTION: Adolescent girls and young women (AGYW) in Nepal have disproportionately poor reproductive and maternal health outcomes. In response, Save the Children, the Nepal government, and local partners designed and implemented Healthy Transitions for Nepali Youth, a multi-level integrated intervention. The intervention aimed to improve reproductive, maternal, and newborn health knowledge, attitudes, and behaviors among AGYW, and address gender attitudes and norms in four districts of Karnali Province, Nepal. METHODS: Married and unmarried AGYW aged 15-24 were engaged in a small group, curriculum-based intervention; husbands and families received home visits, using short videos to catalyze discussion; communities were engaged through dialogue-based activities; and the health system was made more adolescent-responsive through quality assessments, training, and supervision. An external organization conducted a quantitative survey with a sample of 786 AGYW intervention participants at baseline and 565 of the same AGYW at endline. Pooled linear regressions were estimated for each indicator to assess the statistical significance of differences between baseline and endline. Focus group discussions and key informant interviews were conducted with AGYW, husbands, families, community leadership, and program implementers. Data analysis was done through STATA 14th version and NVivo. RESULTS: The percentage of AGYW currently using a modern contraceptive method increased significantly, and more AGYW believed that their family was supportive of delaying marriage and motherhood at the endline. Young women's knowledge of danger signs during labor increased, and there was a significant improvement in essential newborn care practices immediately after birth. AGYW reported shifts towards more gender equitable attitudes and behaviors, including related to decision-making about reproductive and maternal health. CONCLUSION: Positive shifts in reproductive, maternal, and newborn health and gender knowledge, attitudes, and behavior were observed among AGYW, their male partners, and families. The results can inform the design of future interventions to effectively reach this critical population. TRAIL REGISTRATION: Not applicable.

Human dermal fibroblast-derived exosomes induce macrophage activation in systemic sclerosis.

OBJECTIVES: Prior work demonstrates that co-cultured macrophages and fibroblasts from patients with SSc engage in reciprocal activation. However, the mechanism by which these cell types communicate and contribute to fibrosis and inflammation in SSc is unknown. METHODS: Fibroblasts were isolated from skin biopsies obtained from 7 SSc patients or 6 healthy age and gender-matched control subjects following written informed consent. Human donor-derived macrophages were cultured with exosomes isolated from control or SSc fibroblasts for an additional 48 h. Macrophages were immunophenotyped using flow cytometry, qRT-PCR and multiplex. For mutual activation studies, exosome-activated macrophages were co-cultured with SSc or healthy fibroblasts using Transwells. RESULTS: Macrophages activated with dermal fibroblast-derived exosomes from SSc patients upregulated surface expression of CD163, CD206, MHC Class II and CD16 and secreted increased levels of IL-6, IL-10, IL-12p40 and TNF compared with macrophages incubated with healthy control fibroblasts (n = 7, P < 0.05). Exosome-stimulated macrophages and SSc fibroblasts engaged in reciprocal activation, as production of collagen and fibronectin was significantly increased in SSc fibroblasts receiving signals from SSc exosome-stimulated macrophages (n = 7, P < 0.05). CONCLUSION: In this work, we demonstrate for the first time that human SSc dermal fibroblasts mediate macrophage activation through exosomes. Our findings suggest that macrophages and fibroblasts engage in cross-talk in SSc skin, resulting in mutual activation, inflammation, and extracellular matrix (ECM) deposition. Collectively, these studies implicate macrophages and fibroblasts as cooperative mediators of fibrosis in SSc and suggest therapeutic targeting of both cell types may provide maximal benefit in ameliorating disease in SSc patients.

Cystic hepatic metastases in treatment naïve neuroendocrine tumor: Cytomorphology clubbed with cell block immunocytochemistry clinches the diagnosis.

Cystic hepatic lesions encompass a vast spectrum of infectious, non-neoplastic, and neoplastic entities. Most hepatic cysts are benign and asymptomatic, requiring no active intervention. However, symptomatic and malignant cysts need proper evaluation and specific treatment. An accurate preoperative diagnosis is pivotal for patient management. At times, these lesions may mimic, especially symptomatology and radiology, leading to a diagnostic ordeal. We herein present a case of a patient with cystic liver lesions in a treatment naïve neuroendocrine tumor that was a diagnostic dilemma on radiology. Low cellularity of cytology smears made the diagnosis challenging, and cell block immunocytochemistry finally clinched the diagnosis. A timely diagnosis provided on cytology fuelled further work-up, the discovery of primary tumor, and initiation of appropriate therapy.

Unlocking the potential for engaging men to improve reproductive, maternal, and neonatal health in Karnali Province, Nepal.

BACKGROUND: Adolescent girls and young women (AGYW) often experience early childbearing and have poor utilization of reproductive, maternal, and neonatal health (RMNH) services in Nepal. Involving men in such services has been increasingly recognized globally to improve gender-equitable reproductive health behaviour in husbands. This qualitative study assessed the implementation of Healthy Transitions' male engagement interventions in Karnali Province, Nepal which were implemented to improve gender-equitable attitudes, and supportive RMNH care-seeking behaviors among the husbands of young women. METHODS: We conducted a summative qualitative study that included in-depth interviews with 12 AGYW as primary beneficiaries and their husbands (N = 12) and in-laws (N = 8). In addition, key informant interviews were conducted with health workers (N = 8), local government representatives (N = 4), members of Health Facility Operation and Management Committee (N = 8) and project implementers (N = 12). Due to COVID-19-related travel restrictions and lockdowns, all interviews were conducted via phone calls and online consultation. Data were analyzed using multistage coding and thematic content analysis. RESULTS: AGYW, their husbands, in-laws and health workers were receptive to the Healthy transitions' male engagement initiatives. They perceived that the project contributed a momentum to facilitate men's gender-responsive behaviour. Many participants reported that male engagement interventions, including home visits, community dialogues, and social events improved husbands' support for their wives during menstruation, pregnancy, and childbirth. The activities also facilitated spousal communication and improved the couple's decision-making for family planning use. Women reported that improved support from their husbands increased their self-confidence. CONCLUSIONS: This study sheds light on the role of male engagement strategies to improve RMNH in a context where inequitable gender norms and roles are highly prevalent. Our findings highlight the potential to improve RMNH by addressing barriers to male engagement.

Scrub Typhus Presenting as Unilateral Abducens Nerve Palsy: A Case Report.

Abducens nerve palsies associated with infectious diseases are rare. Scrub typhus is an acute, febrile, infectious illness caused by Orientia tsutsugamushi carried out by vector mite zoonosis and is highly endemic in the so-called "tsutsugamushi triangle". The organism has been reported to be capable of entering the nervous system, causing meningitis and focal neurologic abnormalities. We report a 23 years old previously healthy girl who presented with fever, pain abdomen, vomiting and classical pathognomic black eschar mark on the right proximal medial calf region. After exclusion of other common infectious causes, scrub typhus serology immunoglobulin M was positive and was diagnosed with scrub typhus associated with unilateral abducens nerve palsy which responded to doxycycline therapy. On the background of strong clinical suspicion, we underline its significance in the interpretation of the serologic testing and its role in guiding the further treatment respectively. Keywords: abducens nerve palsy; case report; Orientia tsutsugamushi; scrub typhus; zoonoses.

T Cells and CDDO-Me Attenuate Immunosuppressive Activation of Human Melanoma-Conditioned Macrophages.

Melanoma tumors are highly immunogenic, making them an attractive target for immunotherapy. However, many patients do not mount robust clinical responses to targeted therapies, which is attributable, at least in part, to suppression of immune responses by tumor-associated macrophages (TAMs) in the tumor microenvironment (TME). Using a human in vitro tri-culture system of macrophages with activated autologous T cells and BRAFV600E mutant melanoma cells, we now show that activated T cells and the synthetic triterpenoid the methyl ester of 2-cyano-3,12-dioxooleana-1,9(11)-dien-28-oic acid (CDDO-Me) attenuate immune suppression. Surface expression of CD206, CD16 and CD163 on melanoma-conditioned macrophages was inhibited by the addition of T cells, suggesting relief of immuno-suppressive macrophage activation. We also demonstrated that addition of CDDO-Me to tri-cultures enhanced T cell-mediated reductions in CCL2, VEGF and IL-6 production in a contact-independent manner. Because these results suggest CDDO-Me alters melanoma-conditioned macrophage activation, we interrogated CDDO-Me-mediated changes in macrophage signaling pathway activation. Our results indicated that CDDO-Me inhibited phosphorylation of STAT3, a known inducer of TAM activation. Collectively, our studies suggest that activated T cells and CDDO-Me synergistically relieve immune suppression in melanoma cultures and implicate the potential utility of CDDO-Me in the treatment of melanoma.

Self-Assembled Human Skin Equivalents Model Macrophage Activation of Cutaneous Fibrogenesis in Systemic Sclerosis.

OBJECTIVE: The development of precision therapeutics for systemic sclerosis (SSc) has been hindered by the lack of models that accurately mimic the disease in vitro. This study was undertaken to design and test a self-assembled skin equivalent (saSE) system that recapitulates the cross-talk between macrophages and fibroblasts in cutaneous fibrosis. METHODS: SSc-derived dermal fibroblasts (SScDFs) and normal dermal fibroblasts (NDFs) were cultured with CD14+ monocytes from SSc patients or healthy controls to allow de novo stroma formation. Monocyte donor-matched plasma was introduced at week 3 prior to seeding keratinocytes to produce saSE with a stratified epithelium. Tissue was characterized by immunohistochemical staining, atomic force microscopy, enzyme-linked immunosorbent assay, and quantitative reverse transcriptase-polymerase chain reaction. RESULTS: Stroma synthesized de novo from NDFs and SScDFs supported a fully stratified epithelium to form saSE. A thicker and stiffer dermis was generated by saSE with SScDFs, and more interleukin-6 and transforming growth factor ß (TGFß) was secreted by saSE with SScDFs compared to saSE with NDFs, regardless of the inclusion of monocytes. Tissue with SSc monocytes and plasma had amplified dermal thickness and stiffness relative to control tissue. Viable CD163+ macrophages were found within the stroma of saSE 5 weeks after seeding. Additionally, SSc saSE contained greater numbers of CD163+ and CD206+ macrophages compared to control saSE. TGFß blockade inhibited stromal stiffness to a greater extent in SSc saSE compared to control saSE. CONCLUSION: These data suggest reciprocal activation between macrophages and fibroblasts that increases tissue thickness and stiffness, which is dependent in part on TGFß activation. The saSE system may serve as a platform for preclinical therapeutic testing and for molecular characterization of SSc skin pathology through recapitulation of the interactions between macrophages and fibroblasts.

Ecdysone exerts biphasic control of regenerative signaling, coordinating the completion of regeneration with developmental progression.

In Drosophila melanogaster, loss of regenerative capacity in wing imaginal discs coincides with an increase in systemic levels of the steroid hormone ecdysone, a key coordinator of their developmental progression. Regenerating discs release the relaxin hormone Dilp8 (Drosophila insulin-like peptide 8) to limit ecdysone synthesis and extend the regenerative period. Here, we describe how regenerating tissues produce a biphasic response to ecdysone levels: lower concentrations of ecdysone promote local and systemic regenerative signaling, whereas higher concentrations suppress regeneration through the expression of broad splice isoforms. Ecdysone also promotes the expression of wingless during both regeneration and normal development through a distinct regulatory pathway. This dual role for ecdysone explains how regeneration can still be completed successfully in dilp8- mutant larvae: higher ecdysone levels increase the regenerative activity of tissues, allowing regeneration to reach completion in a shorter time. From these observations, we propose that ecdysone hormone signaling functions to coordinate regeneration with developmental progression.

Ecdysone regulates the Drosophila imaginal disc epithelial barrier, determining the length of regeneration checkpoint delay.

Regeneration of Drosophila imaginal discs, larval precursors to adult tissues, activates a regeneration checkpoint that coordinates regenerative growth with developmental progression. This regeneration checkpoint results from the release of the relaxin-family peptide Dilp8 from regenerating imaginal tissues. Secreted Dilp8 protein is detected within the imaginal disc lumen, in which it is separated from its receptor target Lgr3, which is expressed in the brain and prothoracic gland, by the disc epithelial barrier. Here, we demonstrate that following damage the imaginal disc epithelial barrier limits Dilp8 signaling and the duration of regeneration checkpoint delay. We also find that the barrier becomes increasingly impermeable to the transepithelial diffusion of labeled dextran during the second half of the third instar. This change in barrier permeability is driven by the steroid hormone ecdysone and correlates with changes in localization of Coracle, a component of the septate junctions that is required for the late-larval impermeable epithelial barrier. Based on these observations, we propose that the imaginal disc epithelial barrier regulates the duration of the regenerative checkpoint, providing a mechanism by which tissue function can signal the completion of regeneration.

Clinical Profile of COVID-19 Patients Admitted in a COVID Designated Hospital.

BACKGROUND: Pandemic of COVID-19 has engulfed Nepal as well. In this paper, we studied the demographic, clinical, laboratory findings as well as the treatment modalities, prognostic factors and outcome of patients admitted with COVID-19. METHODS: This was an observational cross-sectional study that included all patients admitted to the General Medicine Department of College of Medical Sciences, Bharatpur, during the first wave of COVID-19 from April 2020 to February 2021 after obtaining the ethical clearance. Data analysis was done using statistical packages for social sciences version 16. RESULTS: A total of 119 patients with mean age of 61.5 years were admitted. They had a mean duration of onset of symptoms of 7.1 days. Commonest symptoms were fever (70.6%), cough (67.2%) and dyspnea (64.7%). Severe COVID-19 at admission with a median CT severity score of 15 was found in 49.7% of them. Total 83.2% patients required ICU care and 10.9% required mechanical ventilation. ARDS and secondary infection occurred in 17.6% each. Median length of hospital stay was 6 days. In total, 56.3% recovered 27.7% left against medical advice and 16.0% expired. Severity of COVID at admission, CT severity score at presentation and D-dimer at admission were found to be significantly associated with mortality (P<0.05).Neither of the age, duration of illness, CRP at admission nor the use of remdesivir or convalescent plasma had significant relation with the mortality (P>0.05). CONCLUSIONS: Severity of illness at presentation, CT severity score and D-dimer level at admission are significantly associated with mortality of the patients admitted with COVID-19.

The platelet receptor CLEC-2 blocks neutrophil mediated hepatic recovery in acetaminophen induced acute liver failure.

Acetaminophen (APAP) is the main cause of acute liver failure in the West. Specific efficacious therapies for acute liver failure (ALF) are limited and time-dependent. The mechanisms that drive irreversible acute liver failure remain poorly characterized. Here we report that the recently discovered platelet receptor CLEC-2 (C-type lectin-like receptor) perpetuates and worsens liver damage after toxic liver injury. Our data demonstrate that blocking platelet CLEC-2 signalling enhances liver recovery from acute toxic liver injuries (APAP and carbon tetrachloride) by increasing tumour necrosis factor-α (TNF-α) production which then enhances reparative hepatic neutrophil recruitment. We provide data from humans and mice demonstrating that platelet CLEC-2 influences the hepatic sterile inflammatory response and that this can be manipulated for therapeutic benefit in acute liver injury. Since CLEC-2 mediated platelet activation is independent of major haemostatic pathways, blocking this pathway represents a coagulopathy-sparing, specific and novel therapy in acute liver failure.

CDDO-Me Alters the Tumor Microenvironment in Estrogen Receptor Negative Breast Cancer.

The tumor microenvironment (TME) is an essential contributor to the development and progression of malignancy. Within the TME, tumor associated macrophages (TAMs) mediate angiogenesis, metastasis, and immunosuppression, which inhibits infiltration of tumor-specific cytotoxic CD8+ T cells. In previous work, we demonstrated that the synthetic triterpenoid CDDO-methyl ester (CDDO-Me) converts breast TAMs from a tumor-promoting to a tumor-inhibiting activation state in vitro. We show now that CDDO-Me remodels the breast TME, redirecting TAM activation and T cell tumor infiltration in vivo. We demonstrate that CDDO-Me significantly attenuates IL-10 and VEGF expression but stimulates TNF production, and reduces surface expression of CD206 and CD115, markers of immunosuppressive TAMs. CDDO-Me treatment redirects the TAM transcriptional profile, inducing signaling pathways associated with immune stimulation, and inhibits TAM tumor infiltration, consistent with decreased expression of CCL2. In CDDO-Me-treated mice, both the absolute number and proportion of splenic CD4+ T cells were reduced, while the proportion of CD8+ T cells was significantly increased in both tumors and spleen. Moreover, mice fed CDDO-Me demonstrated significant reductions in numbers of CD4+ Foxp3+ regulatory T cells within tumors. These results demonstrate for the first time that CDDO-Me relieves immunosuppression in the breast TME and unleashes host adaptive anti-tumor immunity.

Profibrotic Activation of Human Macrophages in Systemic Sclerosis.

OBJECTIVE: Genome-wide gene expression studies implicate macrophages as mediators of fibrosis in systemic sclerosis (SSc), but little is known about how these cells contribute to fibrotic activation in SSc. We undertook this study to characterize the activation profile of SSc monocyte-derived macrophages and assessed their interaction with SSc fibroblasts. METHODS: Plasma and peripheral blood mononuclear cells (PBMCs) were obtained from whole blood from SSc patients (n = 24) and age- and sex-matched healthy controls (n = 12). Monocytes were cultured with autologous or allogeneic plasma to differentiate cells into macrophages. For reciprocal activation studies, macrophages were cocultured with fibroblasts using Transwell plates. RESULTS: The gene expression signature associated with blood-derived human SSc macrophages was enriched in SSc skin in an independent cohort and correlated with skin fibrosis. SSc macrophages expressed surface markers associated with activation and released CCL2, interleukin-6, and transforming growth factor ß under basal conditions (n = 8) (P < 0.05). Differentiation of healthy donor monocytes in plasma from SSc patients conferred the immunophenotype of SSc macrophages (n = 13) (P < 0.05). Transwell experiments demonstrated that coculture of SSc macrophages with SSc fibroblasts induced fibroblast activation (n = 3) (P < 0.05). CONCLUSION: These data demonstrate that the activation profile of SSc macrophages is profibrotic. SSc macrophages are activated under basal conditions and release mediators and express surface markers associated with both alternative and inflammatory macrophage activation. These findings also suggest that activation of SSc macrophages arises from soluble factors in local microenvironments. These studies implicate macrophages as likely drivers of fibrosis in SSc and suggest that therapeutic targeting of these cells may be beneficial in ameliorating disease in SSc patients.

High rate of hepatitis C reinfection following antiviral treatment in the North East England Prisons.

To achieve elimination of hepatitis C (HCV), a critical group to prioritise for diagnosis and treatment is the prison population, where HCV prevalence is high. A universal offer of blood-borne virus testing (UOBBVT) programme and a new treatment pathway were introduced to seven North East England (NEE) Prisons. Our aim was to assess: (a) the proportion of individuals with active HCV commencing direct-acting antivirals (DAAs); (b) the outcomes following DAA treatment; (3) the reinfection rate following sustained virological response (SVR). Data were collected prospectively on BBVT uptake, HCV positivity, HCV treatment outcomes and reinfection from March 2016 onwards. 8538 individuals had BBV testing. In total, 612 (7.2%) and 374 (4.4%) were HCV antibody positive and HCV RNA positive, respectively. Ultimately, 266 (71%) individuals commenced DAAs. Overall 111 achieved a documented SVR (42%), 17 (6%) failed treatment, 30 (11%) were still on treatment or had not reached 12 weeks post-treatment at time of analysis, and 108 (41%) were lost to follow-up. In those with a known outcome (n = 128), 87% achieved SVR. Worryingly, of those who achieved SVR, 21 (19%) were subsequently identified as having been reinfected (median time from SVR to documented reinfection 13 (range 7-25) months). The reinfection rate was 0.406 cases per person-year follow-up. In conclusion, Implementation of a UOBBVT programme and new treatment pathway resulted in increased diagnosis and treatment of HCV in the NEE prison population. However, the high HCV reinfection rate suggests a need to improve harm reduction approaches.

Long acting reversible contraception use and associated factors among married women of reproductive age in Nepal.

Modern contraceptives are highly effective and proven means of preventing unintended pregnancy and reducing maternal mortality. Social and economic characteristics are some of the key determinants of health and utilization family planning. However, studies examining the factors associated with utilization of long acting reversible contraception (LARC) are limited in Nepal. This study assessed the factors associated with utilization of LARC methods among married women of reproductive age in Nepal. Secondary data analysis was conducted using the 2016 Nepal Demographic and Health Survey (NDHS). A logistic regression model examined the association of socioeconomic, demographic, or fertility related characteristics with the use of LARCs among 9875 ever married women of reproductive age. The overall utilization rate of LARC in this study was 4.7%. Women in the age group of <25 years (AOR: 0.65, 95% CI: 0.45-0.92) and 25-35 years (AOR: 0.70, 95% CI: 0.56-0.89), having husbands with primary education (AOR:0.71; 95%CI: 0.64-0.84) and no education (AOR: 0.54; 95%CI: 0.38-0.73), belonging to Janajatis (AOR: 0.55; 95%CI: 0.42-0.71) and Newars (AOR: 0.29; 95%CI: 0.19-42), poor wealth quintile (AOR: 0.60; 95% CI: 0.45-0.86) had negative association with LARC use. On the other hand, women having their husband as a skilled worker (AOR: 1.49; 95%CI: 1.10-2), having two or less than two children (AOR: 1.46; 95% CI: 1.15-1.186), and having desire for children in future (AOR: 3.24; 95% CI: 2.29-4.57) had positive association with the use of LARC. In this study, younger women's age, low or no husband's education, from indigenous community such as Janajati and Newer, being in lowest wealth quintile negatively influenced the use of LARC. Conversely, women having her husband as skilled worker, parity less than two, and desire of having future children, positively influenced the use of LARC. The study highlights the need to reach women who were in the lower socioeconomic background to improve LARC use.