Synthetic Nucleic Acid Antigens in Localized Scleroderma.

We investigated the impact of synthetic nucleic acid antigens on the autoantibody profiles in patients with localized scleroderma, an autoimmune skin disease. Anti-DNA antibodies, including double-stranded DNA (dsDNA) and single-stranded DNA (ssDNA), are common among autoimmune diseases, such as systemic lupus erythematosus and localized scleroderma. Based on recent studies, we hypothesized that the sequence of nucleic acid antigens has an impact on the autoimmune reactions in localized scleroderma. To test our hypothesis, we synthesized a panel of DNA and RNA antigens and used them for autoantibody profiling of 70 children with localized scleroderma compared with the healthy controls and patients with pediatric systemic lupus erythematosus (as a disease control). Among the tested antigens, dsD4, which contains the sequence of the human oncogene BRAF, showed a particularly strong presence in localized scleroderma but not systemic lupus erythematosus. Disease activity in patients was significantly associated with dsD4 autoantibody levels. We confirmed this result in vivo by using a bleomycin-induced mouse model of localized scleroderma. When administered intraperitoneally, dsD4 promoted an active polyclonal response in the mouse model. Our study highlights sequence specificity for nucleic acid antigens in localized scleroderma that could potentially lead to developing novel early-stage diagnostic tools.

Defining Collective Priorities: Research and Learning Agendas for Family Planning Across 6 Countries.

Evidence should be the foundation for a well-designed family planning (FP) program, but existing evidence is rarely aligned with and/or synthesized to speak directly to FP programmatic needs. Based on our experience cocreating FP research and learning agendas (FP RLAs) in Côte d'Ivoire, Malawi, Mozambique, Nepal, Niger, and Uganda, we argue that FP RLAs can drive the production of coordinated research that aligns with national priorities.To cocreate FP RLAs, stakeholders across 6 countries conducted desk reviews of 349 documents and 106 key informant interviews, organized consultation meetings in each country to prioritize evidence gaps and generate research and learning questions, and, ultimately, formed 6 FP RLAs comprising 190 unique questions. We outline the process for consensus-driven development of FP RLAs and communicate the results of an analysis of the questions in each FP RLA across 4 technical areas: self-care, equity, high impact practices, and youth. Each question was categorized as a learning versus research question, the former indicating an opportunity to synthesize existing evidence and the latter to conduct new research to answer the question. Themes emerging from the data shed light on shared evidence gaps across the 6 countries. We argue that similarities and differences in the questions in each FP RLA reflect the unique implementation experience and context, as well as each country's placement on the FP S-curve. Early uses of the FP RLAs include informing the development of FP costed implementation plans and FP2030 commitments. FP RLAs have also been discussed in multiple thematic working groups. For FP stakeholders, these FP RLAs represent a consensus-based agenda that can guide the generation and synthesis of evidence to answer each country's most pressing questions, ultimately driving progress toward increasingly evidence-based programming and policy.

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.

Cyclic Citrullinated Peptide Aptamer Treatment Attenuates Collagen-Induced Arthritis.

We describe the study of a novel aptamer-based candidate for treatment of seropositive rheumatoid arthritis. The candidate is a nanoparticle-formulated cyclic citrullinated peptide aptamer, which targets autoantibodies and/or the immune reactions leading to antibody production. Due to its specificity, the peptide aptamer nanoparticles might not interfere with normal immune functions as seen with other disease-modifying antirheumatic drugs. Over a 3-week course of treatment, joint swelling and arthritis score in collagen-induced rats were significantly decreased compared with animals treated with phosphate-buffered saline, unloaded nanoparticles, or nanoparticles with a noncitrullinated control peptide. The reduction in joint swelling was associated with decreased anticitrullinated peptide autoantibody levels in the blood. Treatment with aptamer nanoparticles also increased interleukin-10 levels. The effect seen with the proposed treatment candidate could be mediated by upregulation of anti-inflammatory mediators and decreased levels of anticitrullinated peptide antibodies.

Carbon Nanotubes-Potent Carriers for Targeted Drug Delivery in Rheumatoid Arthritis.

Two types of single-walled carbon nanotubes (SWCNTs), HiPco- and carboxyl-SWCNT, are evaluated as drug carriers for the traditional anti-inflammatory drug methotrexate (MTX) and a small interfering RNA (siRNA) targeting NOTCH1 gene. The nanotubes are solubilized by PEGylation and covalently loaded with MTX. The coupling efficiency (CE%) of MTX is 77-79% for HiPco-SWCNT and 71-83% for carboxyl-SWCNT. siRNA is noncovalently attached to the nanotubes with efficiency of 90-97% for HiPco-SWCNT and 87-98% for carboxyl-SWCNT. Through whole body imaging in the second near-infrared window (NIR-II window, 1000-1700 nm), SWCNTs were found to be selectively accumulated in inflamed joints in a serum transfer mouse model. We further investigated the interactions of the siRNA/MTX loaded nanotubes with human blood and mice bone marrow cells. In human blood, both types of unloaded SWCNTs were associated with B cells, monocytes and neutrophils. Interestingly, loading with MTX suppressed SWCNTs targeting specificity to immune cells, especially B cells; in contrast, loading siRNA alone enhanced the targeting specificity. Loading both MTX and siRNA to carboxyl-SWCNT enhanced targeting specificity to neutrophils and monocytes but not B cells. The targeting specificity of SWCNTs can potentially be adjusted by altering the ratio of MTX and siRNA loaded. The combined results show that carbon nanotubes have the potential for delivery of cargo drugs specifically to immune cells involved in rheumatoid arthritis.

Serological comparison of systemic lupus erythematosus with neuropsychiatric lupus using synthetic nucleic acid antigens.

Neuropsychiatric systemic lupus erythematosus is an autoimmune disorder characterized by an irregular exchange between the central nervous system and the immune system, leading to the outbreak of neurological conditions with possible disabling effects. Although neuropsychiatric systemic lupus erythematosus is the most common expression of lupus condition, it is still poorly understood. In this study, we focus on the development of an advantageous method based on the application of synthetic nucleic acids and protein-based antigen arrays in order to characterize autoreactive antibodies in neuropsychiatric systemic lupus erythematosus. We confirmed the benefits of using synthetic oligonucleotides such as assay reproducibility, elevated affinity and specificity to autoreactive antibodies. We also demonstrated presence of autoantibodies towards three particular synthetic double stranded antigens and verify similarity of antinuclear antibody patterns in ordinary lupus and neuropsychiatric systemic lupus erythematosus.

Antibodies to synthetic citrullinated peptide epitope correlate with disease activity and flares in rheumatoid arthritis.

Rheumatoid arthritis (RA), caused by the abnormal recognition of human joint cells by autoimmune antibodies, remains the world's most prevalent autoimmune disease, with over five million people affected and as much as 4% of the population at risk of RA. To prevent rapid disease development, hormonal and anti-inflammatory therapies require fast and reliable RA diagnosis. However, difficulty in detecting early specific biomarkers for RA means that it is unclear when treatment needs to begin. Here, we combined synthesis of citrullinated peptide epitopes with molecular diagnostics to verify a new specific biomarker for early RA diagnosis and flare prediction. A fibrinogen-derived 21-amino-acid-long citrullinated peptide showed high reactivity toward autoantibodies in RA samples. Additionally, the level of antibodies to this epitope was elevated prior to flares. In contrast, other citrullinated protein variants had lower reactivity and poorer sensitivity to disease activity. In conclusion, fibrinogen-derived epitope E2 subjected to citrullination facilitated a reliable RA diagnosis with a strong correlation to disease activity. This is of a high value for the diagnosis and management of RA patients who respond poorly to treatment.

Combined Assay for Detecting Autoantibodies to Nucleic Acids and Apolipoprotein H in Patients with Systemic Lupus Erythematosus.

The complicated clinical picture and biomolecular pattern of human autoimmune diseases (ADs) make knowledge on their etiology still fragmentary. The diagnostic approaches for ADs require improvement both for clinical and research effort to progress. Synthetic biomolecular antigens find growing applications for diagnosis and investigation of ADs. The main goal of this work is to detect interaction between synthetic antigens and autoantibodies in systemic lupus erythematosus within a combined, high-throughput assay. A panel of synthetic antigens has been prepared from DNA, RNA, locked nucleic acids and apolipoprotein H. The binding of synthetic antigens to autoantibodies has been confirmed in sera samples from those with active systemic lupus erythematosus (SLE) by indirect enzyme linked immunosorbent assay. Our study provides an efficient methodology for combined autoantibody profiling in SLE.

Citrullinated Peptide Epitope Targets Therapeutic Nanoparticles to Human Neutrophils.

Multiple drugs have been proposed for reducing harsh symptoms of human rheumatic diseases. However, a targeted therapy with mild to no side effects is still missing. In this study, we have prepared and tested a series of therapeutic nanoparticles for specific targeting of human neutrophils associated with rheumatoid arthritis. In doing this, a series of citrullinated peptide epitopes derived from human proteins, fibrinogen, vimentin, and histone 3, were screened with regard to specific recognition of neutrophils. The most potent epitope proved to be a mutated fragment of an alpha chain in human fibrinogen. Next, a straightforward synthetic strategy was developed for nanoparticles decorated with this citrullinated peptide epitope and an antisense oligonucleotide targeting disease associated microRNA miR-125b-5p. Our study shows that the nanoparticles specifically recognize neutrophils and knock down miR-125b-5p, with no apparent toxicity to human cells. In contrast to organic dendrimers, chitosan-hyaluronic acid formulations do not activate human innate immune response. Our data proves that the strategy we report herein is effective in developing peptide epitopes for decorating delivery vehicles bearing biological drugs, targeted to a specific cell type.

Autoantibodies in Morphea: An Update.

Skin autoimmune conditions belong to a larger group of connective tissue diseases and primarily affect the skin, but might also involve underlying tissues, such as fat tissue, muscle, and bone. Autoimmune antibodies (autoantibodies) play a role in autoimmune skin diseases, such as localized scleroderma also termed morphea, and systemic scleroderma, also called systemic sclerosis (SSc). The detailed studies on the biological role of autoantibodies in autoimmune skin diseases are limited. This results in a few available tools for effective diagnosis and management of autoimmune skin diseases. This review aims to provide an update on the detection and most recent research on autoantibodies in morphea. Several recent studies have indicated the association of autoantibody profiles with disease subtypes, damage extent, and relapse potential, opening up exciting new possibilities for personalized disease management. We discuss the role of existing autoantibody tests in morphea management and the most recent studies on morphea pathogenesis. We also provide an update on novel autoantibody biomarkers for the diagnosis and study of morphea.

Determinants of pregnancy in adolescents in Nepal.

OBJECTIVE: Social factors and prevalent norms in the community determine the proportion of teenage pregnancy in the community. In the light of high risk associated with teenage pregnancy, the socio-cultural determinants, which influence the conception among teenagers in Nepal, need to be understood. These determinants may be modified by suitable interventions to reduce teenage pregnancy. Aim of this study was to examine the socio-cultural determinants of teenage pregnancy in eastern Nepal. METHODS: A case-control study design was selected for comparing the education, economic status, family support and freedom towards conception among teenagers as compared to higher age group women. RESULT: Seventy adolescent pregnant women were compared with seventy primigravida women in the 20 to 29 years age group. The teenage pregnant women were less educated, had poor economic background, more likely to have accidental pregnancies as compared to the other group and more likely to have love marriages. Husbands were more likely to decide about continuation of pregnancy. They had less psychological and social support from the family. CONCLUSION: Marriage at young age and pregnancy during teens are associated with less social acceptance and poor support in the family.