Awareness and utilization of mother and child protection (MCP) card among mothers attending immunization OPD: An urban health centre-based cross-sectional study.

Background: The MCP card is used as a tool for pregnant women, young mothers and family members to learn about various types of services which they should assess and utilize. The study was undertaken to see the awareness about MCP card and its components among beneficiaries in urban slum area. Methodology: This is a cross-sectional observational study done in urban slum immunization OPD with a sample size of 226. Convenient sampling method was used to select the samples. Mothers consenting and willing to participate in the study were enrolled. Results: Observed that the MCP card was issued to majority participants at the time of baby immunization (44.15%), followed by at time of delivery (31.4%) and least number received it at the time of pregnancy registration (24.34%). It is also observed that only 53.54%(121) women had read the card while the remaining didn't. Majority thought the primary use of MCP card was vaccination record (91.5%), followed by information tool (52.%). It is observed that knowledge regarding pregnancy care and its related complications leads to better utilization of ANC services. Conclusion: As observed in the present study a sizeable proportion of populations doesn't even read the card, and among those who read it the level of knowledge isn't satisfactory, many use the card as a vaccination record but still the awareness about various vaccines among the beneficiaries remains low.

Rubella immunity among pregnant women and the burden of congenital rubella syndrome (CRS) in India, 2022.

BACKGROUND: India aims to eliminate rubella and congenital rubella syndrome (CRS) by 2023. We conducted serosurveys among pregnant women to monitor the trend of rubella immunity and estimate the CRS burden in India following a nationwide measles and rubella vaccination campaign. METHODS: We surveyed pregnant women at 13 sentinel sites across India from Aug to Oct 2022 to estimate seroprevalence of rubella IgG antibodies. Using age-specific seroprevalence data from serosurveys conducted during 2017/2019 (prior to and during the vaccination campaign) and 2022 surveys (after the vaccination campaign), we developed force of infection (FOI) models and estimated incidence and burden of CRS. RESULTS: In 2022, rubella seroprevalence was 85.2% (95% CI: 84.0, 86.2). Among 10 sites which participated in both rounds of serosurveys, the seroprevalence was not different between the two periods (pooled prevalence during 2017/2019: 83.5%, 95% CI: 82.1, 84.8; prevalence during 2022: 85.1%, 95% CI: 83.8, 86.3). The estimated annual incidence of CRS during 2017/2019 in India was 218.3 (95% CI: 209.7, 226.5) per 100, 000 livebirths, resulting in 47,120 (95% CI: 45,260, 48,875) cases of CRS every year. After measles-rubella (MR) vaccination campaign, the estimated incidence of CRS declined to 5.3 (95% CI: 0, 21.2) per 100,000 livebirths, resulting in 1141 (95% CI: 0, 4,569) cases of CRS during the post MR-vaccination campaign period. CONCLUSION: The incidence of CRS in India has substantially decreased following the nationwide MR vaccination campaign. About 15% of women in childbearing age in India lack immunity to rubella and hence susceptible to rubella infection. Since there are no routine rubella vaccination opportunities for this age group under the national immunization program, it is imperative to maintain high rates of rubella vaccination among children to prevent rubella virus exposure among women of childbearing age susceptible for rubella.

Safety and efficacy of danazol in immune thrombocytopenia: a systematic review.

Background: The treatment landscape for relapsed or refractory immune thrombocytopenia (ITP) after corticosteroids is complex. Objectives: We aimed to assess the efficacy of danazol in treating ITP and evaluate the safety and adverse events following its administration. Methods: We searched the databases PubMed, EMBASE, and ClinicalTrials.gov for all published studies assessing danazol's efficacy and safety in treating ITP. The retrieved studies were screened by title and abstract, followed by full-text screening based on the eligibility requirements. The quality assessment was performed using a set of questionnaires. The data were extracted on the descriptive characteristics of the studies and participants, drug dosage, efficacy measures, and adverse effects, and the data were synthesized. Results: A total of 17 studies consisting of 901 participants were included. The overall response rate is around 61% in this analysis. Among the participants, 315 (34.9%) were men. The age of participants ranged from 16 to 86 years. Danazol combined with other pharmacologic interventions, including all-trans-retinoic acid or glucocorticoids, generated better results. The most common side effects appear to be liver injury and elevation of liver enzymes, weight gain, oligomenorrhea, amenorrhea, and myalgia. Conclusion: Danazol at low-to-medium doses was well tolerated and succeeded in improving ITP. Danazol therapy may be helpful in the treatment of chronic ITP that is corticosteroid refractory and when corticosteroids or splenectomy (or both) is contraindicated. Danazol can be considered for further research and development in treating primary immune thrombocytopenia.

Levels of Health Literacy and Associated Factors Among Adults in a Rural Municipality of Nepal: A Cross-sectional Study.

Health literacy is crucial in maintaining good health and well-being, yet community-based studies on health literacy in Nepal are scarce. This study aimed to assess the level of health literacy and identify associated factors among adults in Rasuwa, Nepal. A community-based cross-sectional study was conducted among 253 adults aged above 18 years residing in Kalika Rural Municipality, Rasuwa. Data were collected through face-to-face interviews using the Psychometric Assessment of the European Health Literacy Survey Questionnaire (HLS-EU-Q16). Multivariable logistic regression was used to evaluate independent correlates of health literacy levels. The mean ± SD age of participants was 46.3 ± 16 years. Only about a quarter (23.3%) of participants had adequate health literacy. The participants aged ≤ 45 years (aOR:1.9,95% CI: 1.0-3.6), people who perceived their health status as satisfactory (aOR: 3.1,95% CI: 1.5-6.3), had satisfactory self-rated financial status (aOR: 2.9,95% CI: 1.5-5.5), had satisfactory level of self- related esteem (aOR = 2.7,95% CI: 1.2-6.2), had a stable income (aOR = 1.9, 95% CI:1.0-3.5) were more likely to demonstrate adequate health literacy level. Conversely, illiterate participants (aOR: 0.1, 95% CI: 0.04-0.4) and those engaged in agriculture (aOR: 0.49, 95% CI: 0.2-0.9) had lower odds of having adequate health literacy. This study highlights inadequate health literacy status among approximately three-quarters of the participants. Factors such as age, self-rated health status, financial stability, self-esteem, and income were associated with health literacy levels. These findings underscore the importance of addressing socio-economic and individual factors in promoting health literacy initiatives, particularly targeting vulnerable populations, to improve overall health outcomes and healthcare utilization.

An Unusual Ocular Manifestation of Disseminated Tuberculosis: Combined Retinal Vascular Occlusion Inducing Acute Loss of Vision.

Disseminated tuberculosis (TB) is a life-threatening disease caused by the hematogenous spread of Mycobacterium tuberculosis. Acute loss of vision as a symptom of disseminated TB is uncommon, as per the literature. Uveitis is the most common ocular manifestation of TB, and tubercular retinal arterial or venous occlusion, with or without ocular signs, has been rarely described before. We discuss the case of a 34-year-old truck driver who presented with fever, cough, and sudden painless loss of vision in the right eye. Examination revealed optic neuropathy, as well as central retinal artery and venous occlusion. Investigations showed bilateral miliary shadows on chest X-ray and multiple ring-enhancing brain lesions on MRI brain, consistent with disseminated TB. Anti-tubercular therapy led to clinical improvement. We report this case to highlight the rarity of this condition.

Knowledge, practice, and infection status of prisoners toward dermatomycosis: a study in Nepal.

Background: Dermatophytosis, commonly known as tinea, poses a significant public health concern worldwide, especially in environments with poor hygiene and overcrowding, such as prisons. Despite its prevalence and impact on quality of life, there is a lack of research on the knowledge and hygiene practices regarding dermatophytosis among prisoners, particularly in Nepal. Objective: The study aimed to assess prisoners' knowledge, hygiene practice and infection status regarding dermatophytosis in Central Prison, Nepal. Methods: A descriptive cross-sectional study with a sample size of 184 respondents was designed to collect data using a validated pre-tested questionnaire from September 2023 to January 2024. The collected data was then analyzed using IBM SPSS version 21. Knowledge and hygiene practices were measured on an eight and 11-point scale and rated as poor (≤4) and sound (>4), bad (≤6), and good (>6), respectively. Summary data were presented by descriptive, while Chi-square and logistic regression were used for inferential statistics at p < 0.05. Results: The findings revealed moderate knowledge among prisoners regarding dermatophytosis, with significant gaps in understanding its spread and prevention. While most prisoners recognized the importance of treatment, there were misconceptions about the inevitability of contracting dermatophytosis and the role of personal hygiene. Conclusion: Despite good knowledge levels, adherence to recommended hygiene practices was suboptimal, highlighting the need for targeted interventions. The study underscores the importance of addressing knowledge gaps, changing attitudes, and promoting hygienic practices to mitigate the burden of dermatophytosis among prisoners.

A baseline cross-sectional study to assess patient satisfaction (18-60 years) toward Apli Chikitsa (free of cost medical laboratory tests) services provided at an urban health training center: A PPP model.

Introduction: Assessing satisfaction with healthcare services is one of the cornerstone methods to evaluate success and identify obstacles during the process for the patients. Identifying crucial information helps in course correction and better healthcare delivery to the patients. Apli Chikitsa is a novel PPP model-based free of cost laboratory service made available to the patients by the municipal corporation. Around 150 blood tests are offered free of cost and through peripheral dispensaries to the patients. The present study was undertaken to assess the satisfaction of patients with the service and service-related aspects. Methodology: A cross-sectional observational urban health center-based study was undertaken to assess the satisfaction of patients toward the Apli Chikitsa laboratory service. Institutional ethics committee clearance was obtained before commencing the study. Four hundred patients availing of the laboratory services were enrolled by alternate sampling method. A semistructured questionnaire was administered; five-point Likert scale was used to assess satisfaction toward the service and related aspects. Results were summarized using tables and charts, and percentages and proportions were used wherever necessary. Data were analyzed using Microsoft Excel. Results: Among the study participants, overall satisfaction was observed among 71.25%, and 100% satisfaction was observed in relation to cost of service, maintenance of privacy and confidentiality, and getting all the requested tests done. The problem of overcrowding (96.75%) was observed by a majority. Satisfaction was low regarding the cleanliness of toilets (48.75%). Conclusion: Based on the findings, it can be concluded that the overall satisfaction is the Apli Chikitsa laboratory service is good, and it is a good model for replication for laboratory service delivery in peripheral areas away from tertiary care centers. Overcrowding should be managed, and cleanliness of toilets to done at frequent intervals, along with proper and visible use of signages, should be done to improve the satisfaction of the patients.

A critical review of therapeutic interventions in sickle cell disease: Progress and challenges.

Sickle cell disease (SCD) is an autosomal recessive genetic disorder that occurs due to the point mutation in the ß-globin gene, which results in the formation of sickle hemoglobin (HbS) in the red blood cells (RBCs). When HbS is exposed to an oxygen-depleted environment, it polymerizes, resulting in hemolysis, vaso-occlusion pain, and impaired blood flow. Still, there is no affordable cure for this inherited disease. Approved medications held promise but were met with challenges due to limited patient tolerance and undesired side effects, thereby inhibiting their ability to enhance the quality of life across various individuals with SCD. Progress has been made in understanding the pathophysiology of SCD during the past few decades, leading to the discovery of novel targets and therapies. However, there is a compelling need for research to discover medications with improved efficacy and reduced side effects. Also, more clinical investigations on various drug combinations with different mechanisms of action are needed. This review comprehensively presents therapeutic approaches for SCD, including those currently available or under investigation. It covers fundamental aspects of the disease, such as epidemiology and pathophysiology, and provides detailed discussions on various disease-modifying agents. Additionally, expert insights are offered on the future development of pharmacotherapy for SCD.

Pontine tubercular abscess: A rare presentation of CNS tuberculosis masquerading as glioma in a child.

Tuberculous brain abscess (TBA) in a child was initially misdiagnosed as glioma. Two craniotomies, abscess drainage, and anti-tubercular therapy led to recovery. Pontine TBA, though rare and atypical, can have better outcome with timely intervention.

Epistasis and pleiotropy-induced variation for plant breeding.

Epistasis refers to nonallelic interaction between genes that cause bias in estimates of genetic parameters for a phenotype with interactions of two or more genes affecting the same trait. Partitioning of epistatic effects allows true estimation of the genetic parameters affecting phenotypes. Multigenic variation plays a central role in the evolution of complex characteristics, among which pleiotropy, where a single gene affects several phenotypic characters, has a large influence. While pleiotropic interactions provide functional specificity, they increase the challenge of gene discovery and functional analysis. Overcoming pleiotropy-based phenotypic trade-offs offers potential for assisting breeding for complex traits. Modelling higher order nonallelic epistatic interaction, pleiotropy and non-pleiotropy-induced variation, and genotype × environment interaction in genomic selection may provide new paths to increase the productivity and stress tolerance for next generation of crop cultivars. Advances in statistical models, software and algorithm developments, and genomic research have facilitated dissecting the nature and extent of pleiotropy and epistasis. We overview emerging approaches to exploit positive (and avoid negative) epistatic and pleiotropic interactions in a plant breeding context, including developing avenues of artificial intelligence, novel exploitation of large-scale genomics and phenomics data, and involvement of genes with minor effects to analyse epistatic interactions and pleiotropic quantitative trait loci, including missing heritability.

Triggers and factors associated with moral distress and moral injury in health and social care workers: A systematic review of qualitative studies.

OBJECTIVE: At some point in their career, many healthcare workers will experience psychological distress associated with being unable to take morally or ethically correct action, as it aligns with their own values; a phenomenon known as moral distress. Similarly, there are increasing reports of healthcare workers experiencing long-term mental and psychological pain, alongside internal dissonance, known as moral injury. This review examined the triggers and factors associated with moral distress and injury in Health and Social Care Workers (HSCW) employed across a range of clinical settings with the aim of understanding how to mitigate the effects of moral distress and identify potential preventative interventions. METHODS: A systematic review was conducted and reported according to recommendations from Cochrane and Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines. Searches were conducted and updated regularly until January 2024 on 2 main databases (CENTRAL, PubMed) and three specialist databases (Scopus, CINAHL, PsycArticles), alongside hand searches of study registration databases and other systematic reviews reference lists. Eligible studies included a HSCW sample, explored moral distress/injury as a main aim, and were written in English or Italian. Verbatim quotes were extracted, and article quality was assessed via the CASP toolkit. Thematic analysis was conducted to identify patterns and arrange codes into themes. Specific factors like culture and diversity were explored, and the effects of exceptional circumstances like the pandemic. RESULTS: Fifty-one reports of 49 studies were included in the review. Causes and triggers were categorised under three domains: individual, social, and organisational. At the individual level, patients' care options, professionals' beliefs, locus of control, task planning, and the ability to make decisions based on experience, were indicated as elements that can cause or trigger moral distress. In addition, and relevant to the CoVID-19 pandemic, was use/access to personal protection resources. The social or relational factors were linked to the responsibility for advocating for and communication with patients and families, and professionals own support network. At organisational levels, hierarchy, regulations, support, workload, culture, and resources (staff and equipment) were identified as elements that can affect professionals' moral comfort. Patients' care, morals/beliefs/standards, advocacy role and culture of context were the most referenced elements. Data on cultural differences and diversity were not sufficient to make assumptions. Lack of resources and rapid policy changes have emerged as key triggers related to the pandemic. This suggests that those responsible for policy decisions should be mindful of the potential impact on staff of sudden and top-down change. CONCLUSION: This review indicates that causes and triggers of moral injury are multifactorial and largely influenced by the context and constraints within which professionals work. Moral distress is linked to the duty and responsibility of care, and professionals' disposition to prioritise the wellbeing of patients. If the organisational values and regulations are in contrast with individuals' beliefs, repercussions on professionals' wellbeing and retention are to be expected. Organisational strategies to mitigate against moral distress, or the longer-term sequalae of moral injury, should address the individual, social, and organisational elements identified in this review.

Multiplex PCR for Early Generation Identification of Tomato Segregants Carrying Ty-2, Ty-3 and Ph-3 Resistance Alleles Against Leaf Curl and Late Blight Diseases.

Deployment of different natural disease resistance alleles is the most sustainable and eco-friendly way for multiple disease management in tomato. Diagnostic molecular markers are indispensible in this effort as they offer early generation identification of resistance alleles in an environment-independent manner. Moreover, optimized multiplex polymerase chain reaction (PCR) for detecting different disease resistance alleles in a single reaction can speed-up the selection process with cost and labour-effectiveness. Here we report the optimized multiplex detection and stacking of leaf curl disease resistance alleles Ty-2 and Ty-3 along with late blight disease resistance allele Ph-3 in tomato genotypes and F2 segregants. The triplex assay could be replaced by a duplex assay (for Ty-2 and Ty-3 resistance alleles) followed by analysis at Ph-3 locus to achieve further cost-effectiveness. We identified two plants in F2 populations derived from the Arka Samrat (F1) x Kashi Chayan combination to carry the Ty-2, Ty-3 and Ph-3 resistance alleles in homozygous condition. Early generation genotyping also allowed us to identify a few morphologically better segregants, where further marker assisted selection (MAS) should identify superior multiple disease resistant lines. Thus we advocate the utility of multiplex PCR in MAS to address multiple disease resistance breeding in tomato.

Cardiac complications (arrhythmias and heart failure) in patients with ischemic stroke: A meta-analysis.

BACKGROUND: In patients with ischemic stroke (pwIS), cardiac complications have been observed in observational studies. We conducted a systematic review and meta-analysis to investigate the arrhythmias and heart failure in pwIS. METHODS: Up until September 2023, we searched for case-control, cross-sectional, or cohort studies in 4 databases. For case-control/cross-sectional studies, odds ratios (OR) were determined using a random-effects model meta-analysis, while hazard ratios (HR) were calculated for cohort studies, and 95% confidence intervals (CIs) were pooled in the meta-analysis. RESULTS: In the meta-analysis, we incorporated 5 studies: 2 cohort studies, 2 case-control studies, and 1 cross-sectional study. In all, 81,181 controls and 25,544 pwIS were included in this investigation. The combined OR for case-control studies of arrhythmias was estimated to be 1.86 (95% CI: 0.70-4.94, P = .21), HR for cohort studies of arrhythmias to be 4.2 (95% CI: 1.49-12.01, P < .05), and for cohort studies of heart failure to be 2.9 (95% CI: 2.65-3.18, P < .05), suggesting that pwIS may be more likely to experience cardiac complications. CONCLUSION: Our results revealed that there is a comparatively higher risk of cardiac complications in pwIS; however, more research is needed to evaluate the risk of cardiac complications in pwIS.

Efficacy and outcomes of bempedoic acid versus placebo in patients with hypercholesterolemia: an updated systematic review and meta-analysis of randomized controlled trials.

Introduction: Bempedoic acid (BA) has shown varied efficacy in managing hyperlipidemia. We conducted the most extensive up-to-date meta-analysis, the first to include recent studies by Nissen et al., which boast the largest sample size. Methods: Literature search was done on Medline, EMBASE, and Cochrane Library. The primary endpoint was a change in low-density lipoprotein-cholesterol (LDL-C) levels, while secondary endpoints encompassed changes in lipid parameters, clinical endpoints, and safety endpoints. The least-square mean (LSM) percent change was utilized for lipid changes, with statistical significance set at P < 0.05. Results: This analysis included 12 randomized control trials with 22,249 participants. BA exhibited a substantial reduction in LDL-C levels [LSM % change, -24.34; 95% confidence interval (CI), -27.80 to -20.88; P < 0.0001], total cholesterol levels (LSM % change, -16.62; 95% CI, -21.70 to -11.54; P < 0.00001) and high-density lipoprotein-cholesterol (HDL-C) levels (LSM % change, -4.22; 95% CI, -5.51 to -2.92; P < 0.00001) compared to the placebo. Conclusions: BA significantly lowers LDL-C, total cholesterol, HDL-C, non-HDL-C, high sensitivity C reactive protein, and apolipoprotein levels.

Cerebral microbleeds: Causes, clinical relevance, and imaging approach - A narrative review.

With advances in magnetic resonance imaging (MRI) sequences, there has been increased identification of microbleed/microhemorrhage across different population ages, but more commonly in the older age group. These are defined as focal areas of signal loss on gradient echo MRI sequences (T2* and susceptibility-weighted images), which are usually <5 mm in size representing hemosiderin deposition with wide ranges of etiologies. Susceptibility-weighted imaging (SWI) has become a routine MRI sequence for practices across the globe resulting in better identification of these entities. Over the past decade, there has been a better understanding of the clinical significance of microbleeds including their prognostic value in ischemic and hemorrhagic stroke. Cerebral amyloid angiopathy and hypertension are the two most common causes of microbleeds following peripheral and central pattern, respectively. In the younger age group, microbleeds are more common due to familial conditions or a wide range of hypercoagulable states. This review outlines the pathophysiology, prevalence, and clinical implications of cerebral microhemorrhage along with a brief discussion about the technical considerations of SWI.

Impact of a Short Educational Session on Early Diagnosis and Management of Acute Kidney Injury for Different Specialties.

AIM AND OBJECTIVE: This questionnaire study aimed to evaluate the impact of a short educational session on the early diagnosis and management of acute kidney injury (AKI) among doctors specializing in fields other than nephrology, assessed through pre- and post-test scores. This educational study included resident doctors from various specialties for assessment. MATERIALS AND METHODS: The study enrolled different specialty resident doctors' departments and assessed them through questionnaires and assessment scores. The pre-test questionnaires were first distributed and collected after 20 minutes. This was followed by a 30-minute short educational lecture on AKI by the nephrology faculty about its early diagnosis and management as per the Kidney Disease Improving Global Outcomes guidelines. Immediately post continuing medical education, the same questionnaires were distributed along with feedback forms and collected after 10 minutes. RESULTS: A total of 110 residents participated in the study. All participants showed significant improvement in the post-lecture questionnaires compared to pre-lecture scores. For medicine and allied branch residents, the pre- and post-lecture scores were significantly higher than those of the surgical and allied branch residents. The lowest score was observed in residents of orthopedics. The improvement scores of all departments also showed significant differences. The highest improvement was seen in the department of gynecology, followed by residents of the critical care unit and the department of anesthesia. The residents of those departments with high pre- and post-test scores had lower improvement scores. CONCLUSION: The study found a significant knowledge gap in different sister specialties in diagnosing and managing AKI. Short educational sessions showed significant improvement in AKI understanding by addressing the knowledge gaps.

CAIM: Coverage-based Analysis for Identification of Microbiome.

Accurate taxonomic profiling of microbial taxa in a metagenomic sample is vital to gain insights into microbial ecology. Recent advancements in sequencing technologies have contributed tremendously toward understanding these microbes at species resolution through a whole shotgun metagenomic (WMS) approach. In this study, we developed a new bioinformatics tool, CAIM, for accurate taxonomic classification and quantification within both long- and short-read metagenomic samples using an alignment-based method. CAIM depends on two different containment techniques to identify species in metagenomic samples using their genome coverage information to filter out false positives rather than the traditional approach of relative abundance. In addition, we propose a nucleotide-count based abundance estimation, which yield lesser root mean square error than the traditional read-count approach. We evaluated the performance of CAIM on 28 metagenomic mock communities and 2 synthetic datasets by comparing it with other top-performing tools. CAIM maintained a consitently good performance across datasets in identifying microbial taxa and in estimating relative abundances than other tools. CAIM was then applied to a real dataset sequenced on both Nanopore (with and without amplification) and Illumina sequencing platforms and found high similality of taxonomic profiles between the sequencing platforms. Lastly, CAIM was applied to fecal shotgun metagenomic datasets of 232 colorectal cancer patients and 229 controls obtained from 4 different countries and primary 44 liver cancer patients and 76 controls. The predictive performance of models using the genome-coverage cutoff was better than those using the relative-abundance cutoffs in discriminating colorectal cancer and primary liver cancer patients from healthy controls with a highly confident species markers.

cRGD-modified hybrid lipopolymeric nanoplexes for gene editing in the posterior segment of the eye.

Eye-related diseases, specifically retinal dystrophy (RD) conditions, are the leading cause of blindness worldwide. Gene addition, regulation, or editing could potentially treat such diseases through gene expression regulation. CRISPR/Cas9 gene editing is one of the most prominent and precise gene editing tools which could be employed to edit genes related to the dystrophic condition. However, CRISPR/Cas9 faces in vivo delivery challenges due to its high molecular weight, negative charge, prone to degradation in the presence of nucleases and proteases, poor cellular degradation, etc., which makes it challenging to adopt for therapeutic applications. We developed cRGD-modified lipopolymeric nanoplexes loaded with Cas9 RNPs with a particle size and zeta potential of 175 ±â€¯20 nm and 2.15 ±â€¯0.9 mV, respectively. The cRGD-modified lipopolymeric nanoplexes were stable for 194 h and able to transfect >70 % ARPE-19 and NIH3T3 cells with an Indel frequency of ~40 % for the VEGF-A gene. The cRGD-modified lipopolymeric nanoplexes found good vitreous mobility and could transfection retinal cells in vivo after 48 h of intravitreal injection in Wistar Rats. Moreover, in vivo VEGFA gene editing was ~10 % with minimal toxicities. Collectively, the cRGD-modified lipopolymeric nanoplexes were found to have extreme potential in delivering CRISPR/Cas9 RNPs payload to the retinal tissues for therapeutic applications.

Deficiency of the Deubiquitinase UCHL1 Attenuates Pulmonary Arterial Hypertension.

BACKGROUND: The ubiquitin-proteasome system regulates protein degradation and the development of pulmonary arterial hypertension (PAH), but knowledge about the role of deubiquitinating enzymes in this process is limited. UCHL1 (ubiquitin carboxyl-terminal hydrolase 1), a deubiquitinase, has been shown to reduce AKT1 (AKT serine/threonine kinase 1) degradation, resulting in higher levels. Given that AKT1 is pathological in pulmonary hypertension, we hypothesized that UCHL1 deficiency attenuates PAH development by means of reductions in AKT1. METHODS: Tissues from animal pulmonary hypertension models as well as human pulmonary artery endothelial cells from patients with PAH exhibited increased vascular UCHL1 staining and protein expression. Exposure to LDN57444, a UCHL1-specific inhibitor, reduced human pulmonary artery endothelial cell and smooth muscle cell proliferation. Across 3 preclinical PAH models, LDN57444-exposed animals, Uchl1 knockout rats (Uchl1-/-), and conditional Uchl1 knockout mice (Tie2Cre-Uchl1fl/fl) demonstrated reduced right ventricular hypertrophy, right ventricular systolic pressures, and obliterative vascular remodeling. Lungs and pulmonary artery endothelial cells isolated from Uchl1-/- animals exhibited reduced total and activated Akt with increased ubiquitinated Akt levels. UCHL1-silenced human pulmonary artery endothelial cells displayed reduced lysine(K)63-linked and increased K48-linked AKT1 levels. RESULTS: Supporting experimental data, we found that rs9321, a variant in a GC-enriched region of the UCHL1 gene, is associated with reduced methylation (n=5133), increased UCHL1 gene expression in lungs (n=815), and reduced cardiac index in patients (n=796). In addition, Gadd45α (an established demethylating gene) knockout mice (Gadd45α-/-) exhibited reduced lung vascular UCHL1 and AKT1 expression along with attenuated hypoxic pulmonary hypertension. CONCLUSIONS: Our findings suggest that UCHL1 deficiency results in PAH attenuation by means of reduced AKT1, highlighting a novel therapeutic pathway in PAH.