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Leila Gobejishvili,1 Walter E Rodriguez,1 Philip Bauer,2 Yali Wang,1 Chirag Soni,2 Todd Lydic,3 Shirish Barve,1 Craig McClain,1 Claudio Maldonado4 1Department of Medicine, School of Medicine, University of Louisville, Louisville, KY, USA; 2Endoprotech, Inc., Louisville, KY, USA; 3Lipidomics Center, Michigan State University, East Lansing, MI, USA; 4Department of Physiology, School of Medicine, University of Louisville, Louisville, KY, USACorrespondence: Claudio Maldonado, Department of Physiology, School of Medicine, University of Louisville, 500 S. Preston Street, HSC A-1115, Louisville, KY, 40292, USA, Tel +1 (502) 852-1078, Email claudio.maldonado@louisville.edu Leila Gobejishvili, Department of Medicine, School of Medicine, University of Louisville, 505 S. Hancock Street, CTR 516, Louisville, KY, 40202, USA, Tel +1 (502) 852-0361, Fax +1 (502) 852-8927, Email l0gobe01@louisville.eduIntroduction: The phosphodiesterase 4 (PDE4) inhibitor, rolipram, has beneficial effects on tissue inflammation, injury and fibrosis, including in the liver. Since rolipram elicits significant CNS side-effects in humans (ie, nausea and emesis), our group developed a fusogenic lipid vesicle (FLV) drug delivery system that targets the liver to avoid adverse events. We evaluated whether this novel liposomal rolipram formulation reduces emesis.Methods: C57Bl/6J male mice were used to compare the effect of three doses of free and FLV-delivered (FLVs-Rol) rolipram in a behavioral correlate model of rolipram-induced emesis. Tissue rolipram and rolipram metabolite levels were measured using LC-MS/MS. The effect of FLVs-Rol on brain and liver PDE4 activities was evaluated.Results: Low and moderate doses of free rolipram significantly reduced anesthesia duration, while the same doses of FLVs-Rol had no effect. However, the onset and duration of adverse effects (shortening of anesthesia period) elicited by a high dose of rolipram was not ameliorated by FLVs-Rol. Post-mortem analysis of brain and liver tissues demonstrated that FLVs affected the rate of rolipram uptake by liver and brain. Lastly, administration of a moderate dose of FLVs-Rol attenuated endotoxin induced PDE4 activity in the liver with negligible effect on the brain.Discussion: The findings that the low and moderate doses of FLVs-Rol did not shorten the anesthesia duration time suggest that FLV delivery prevented critical levels of drug from crossing the blood-brain barrier (BBB) to elicit CNS side-effects. However, the inability of high dose FLVs-Rol to prevent CNS side-effects indicates that there was sufficient unencapsulated rolipram to cross the BBB and shorten anesthesia duration. Notably, a moderate dose of FLVs-Rol was able to decrease PDE4 activity in the liver without affecting the brain. Taken together, FLVs-Rol has a strong potential for clinical application for the treatment of liver disease without side effects.Keywords: liposomes, rolipram, side-effects, PDE4, liver

A 41-year-old man was referred to our outpatient department with a diagnosis of urticaria with angioedema of 3 months duration. On examination, he had generalized coalescent waxy papules and diffuse periorbital swelling. Systemic examination was unremarkable except for limited finger flexion. Serum electrophoresis and thyroid function tests were normal. Histopathological examination showed normal epidermis and intradermal mucin deposition, which was diagnostic of lichen myxedematosus (LM). The patient showed prompt response to melphalan. Here, we report this case of atypical LM because the patient had generalized eruption with normal thyroid function along with the absence of monoclonal gammopathy.

Background . Fecalomas are hard dense masses separate from surrounding fecal material or bowel contents. This case report intends to provide a brief review of the literature and differential diagnosis for a pelvic mass in a pediatric patient. Case Presentation . The patient is a 5-year-old male presenting with worsening constipation and stool leakage over several months, found to have a rare calcified pelvic mass on abdominal X-ray consistent with a fecaloma. Conclusion . Fecalomas should be considered on the differential diagnosis of pediatric patients who present with chronic constipation and a calcified pelvic mass.

The increasing use of serial PET/CT scans in the management of pediatric malignancies raises the important consideration of radiation exposure in children.To estimate the cumulative radiation dose from PET/CT studies to children with malignancy and to compare with the data in literature.Two hundred forty-eight clinical PET/CT studies performed on 78 patients (50 boys/28 girls, 1.3 to 18 years old from December 2002 to October 2007) were retrospectively reviewed under IRB approval. The whole-body effective dose (ED) estimates for each child were obtained by estimating the effective dose from each PET/CT exam performed using the ImPACT Patient Dosimetry Calculator for CT and OLINDA for PET.The average number of PET/CT studies was 3.2 per child (range: 1 to 14 studies). The average ED of an individual CT study was 20.3 mSv (range: 2.7 to 54.2), of PET study was 4.6 mSv (range: 0.4 to 7.7) and of PET/CT study was 24.8 mSv (range: 6.2 to 60.7). The average cumulative radiation dose per patient from CT studies was 64.4 mSv (range: 2.7 to 326), from PET studies was 14.5 mSv (range: 2.8 to 73) and from PET/CT studies was 78.9 mSv (range: 6.2 to 399).The radiation exposure from serial PET/CT studies performed in pediatric malignancies was considerable; however, lower doses can be used for both PET and CT studies. The ALARA principle must be applied without sacrificing diagnostic information.

Introduction: A prescription audit is a quality improvement procedure that aims to enhance patient care and is a component of the comprehensive clinical audit. Aim: This study was conducted to audit the quality of outpatient department [OPD] prescriptions at an urban government hospital and to analyse the gap between the medicines prescribed and the effective utilization of alternative preparations available in the hospital and the resulting cost burden on the patients. Material and Methods: An observational study was carried out in a government hospital in Pune wherein 1000 prescriptions were collected randomly from patients visiting the hospital pharmacy irrespective of diagnosis over a period of 10 days in January 2023 and May 2023. The prescriptions were analyzed for medical components and drug use indicators and instances where NSAIDS, multivitamins, antidyspeptics and brand names were prescribed along with their dosing. The data obtained was summed up and analyzed using MS-Excel. Results: Around 36.3% prescriptions had diagnosis mentioned in them, 77.9% prescriptions had ≥ 3 drugs, 49% had fixed dose combinations (FDCs), 43.4% had multivitamins, while 51.8% had antidyspeptics. NSAIDs and paracetamol were prescribed to 45.10% patients. Ten percent prescriptions had drugs that were prescribed from outside, but their substitute was available in the hospital pharmacy. Conclusion: Our study highlights the need of scrutiny of prescriptions by senior consultants in terms of dosing and brand names so that drugs can be utilized appropriately. Sharing of inventory with prescribers and orientation with regard to use of available substitutes may be helpful in reducing out of pocket expenditure. Frequently prescribed drugs can be added in the inventory to reduce the cost burden on patients.

Background: Nurses frequently experience compassion fatigue and burnout, which impact their personal lives and patient care. The COVID-19 pandemic additionally caused stress, uncertainty, and fear of death among healthcare professionals. Aim: To assess professional quality of life (ProQoL) among nurses after the second wave of the COVID-19 pandemic. Material and Methods: A web-based cross-sectional study was conducted among 203 nurses using a purposive sampling technique in the month of September to December 2021. Data were collected using a self-administered ProQoL scale version 5. Statistical Analysis: Descriptive and inferential statistics were used. Kolmogorov–Smirnov test, Mann–Whitney U, and Kruskal–Wallis H test were used. Bivariate correlations were used to correlate the main variables. Multiple linear regression analysis was also performed. Results: The majority of the nurses reported a moderate level of compassion satisfaction (CS) (62.6%), burnout (BO) (66.0%), and secondary traumatic stress (STS) (63.1%). Residence and education emerged as a factor whether the nurses experienced BO or STS, respectively. Additionally, CS negatively correlated with BO (r = -0.732: P < 0.001) and STS (r = -0.141: p-0.04). Conclusions: The majority of the nurses experienced moderate levels of CS, BO, and STS after the second wave of the COVID-19 crisis and nurse patient–ratio emerged as a significant factor to predict CS, BO, or STS. Hence, effective measures need to be implemented by hospital administration to enhance the nurses' satisfaction and reduce fatigue and burnout.

Objective: The study aimed to assess the effectiveness of lukewarm water compress on breast pain and breast engorgement among post-cesarean primiparous mothers. Materials and Methods: This quasi-experimental posttest-only design was conducted with 60 post-cesarean section primiparous mothers between October 2018 and January 2019. A simple random sampling technique was used to allocate 30 participants to both experimental and control groups. The experimental group received lukewarm water compress using a sponge cloth for 20 min twice a day on the second, third, and fourth postnatal days. The control group received routine hospital care. Breast pain and engorgement were assessed using the Visual Analog Pain Scale and Breast Engorgement Assessment Scale on the third, fourth, and fifth postnatal days. Results: Significant differences were observed between the experimental and control groups on day 1 and day 3 in terms of breast pain and engorgement scores among post-cesarean section mothers (P < 0.001). In addition, no statistically significant differences were found between sociodemographic and breastfeeding parameters and breast pain and engorgement (P > 0.05). Conclusion: Lukewarm water compress is effective in reducing breast pain and engorgement in post-cesarean primiparous mothers. Future research can include randomized controlled trials to evaluate the effectiveness of different therapies in treating breast pain and engorgement.

Abstract Three gene families in plants viz. Argonaute (AGOs), Dicer-like (DCLs) and RNA dependent RNA polymerase (RDRs) constitute the core components of small RNA mediated gene silencing machinery. The present study endeavours to identify members of these gene families in tea and to investigate their expression patterns in different tissues and various stress regimes. Using genome-wide analysis, we have identified 18 AGOs, 5 DCLs and 9 RDRs in tea, and analyzed their phylogenetic relationship with orthologs of Arabidopsis thaliana. Gene expression analysis revealed constitutive expression of CsAGO1 in all the studied tissues and stress conditions, whereas CsAGO10c showed most variable expression among all the genes. CsAGO10c gene was found to be upregulated in tissues undergoing high meristematic activity such as buds and roots, as well as in Exobasidium vexans infected samples. CsRDR2 and two paralogs of CsAGO4, which are known to participate in biogenesis of hc-siRNAs, showed similarities in their expression levels in most of the tea plant tissues. This report provides first ever insight into the important gene families involved in biogenesis of small RNAs in tea. The comprehensive knowledge of these small RNA biogenesis purveyors can be utilized for tea crop improvement aimed at stress tolerance and quality enhancement.

A 41-year-old man was referred to our outpatient department with a diagnosis of urticaria with angioedema of 3 months duration. On examination, he had generalized coalescent waxy papules and diffuse periorbital swelling. Systemic examination was unremarkable except for limited finger flexion. Serum electrophoresis and thyroid function tests were normal. Histopathological examination showed normal epidermis and intradermal mucin deposition, which was diagnostic of lichen myxedematosus (LM). The patient showed prompt response to melphalan. Here, we report this case of atypical LM because the patient had generalized eruption with normal thyroid function along with the absence of monoclonal gammopathy. [ABSTRACT FROM AUTHOR]

The aim of present study was to prepare novel modified release press coated tablets of venlafaxine hydrochloride. Hydroxypropylmethylcenulose K4M and hydroxypropylmethylcellulose K100M were used as release modifier in core and coat, respectively. A 3² full factorial design was adopted in the optimization study. The drug to polymer ratio in core and coat were chosen as independent variables. The drug release in the first hour and drug release rate between 1 and 12 h were chosen as dependent variables. The tablets were characterized for dimension analysis, crushing strength, friability and in vitro drug release. A check point batch, containing 1:2.6 and 1:5.4 drug to polymer in core and coat respectively, was prepared. The tablets of check point batch were subjected to in vitro drug release in dissolution media with pH 5, 7.2 and distilled water. The kinetics of drug release was best explained by Korsmeyer and Peppas model (anomalous non-Fickian diffusion). The systematic formulation approach enabled us to develop modified release venlafaxine hydrochloride tablets. [ABSTRACT FROM AUTHOR]

Background . Idiopathic intracranial hypertension (IIH), also known as pseudotumor cerebri, is a diagnosis of exclusion in the clinical scenario of increased intracranial pressure without an identifiable cause. This case report provides a brief review of current literature on IIH, important differential considerations to rule out prior to diagnosis, and relevant imaging findings of IIH. Case Presentation . An 18-year-old male presenting with headache and blurry vision was found to have signs of increased intracranial pressure on imaging without other abnormalities to explain the cause, suggesting IIH as a diagnosis. Conclusion . IIH is classically seen in overweight females of childbearing age but should be considered in all obese patients, including the pediatric population given the increasing rate of childhood obesity.

The article presents a clinical image findings of a case involving a 34-weeks gestation premature baby with congenital persistent terminal ventricle and filar cyst. It presents the clinical photos obtained from the baby who has been considered to have a sallow patch on the sacrum raising suspicion of spinal dysraphism. Explanation of the images is also included, with further recommendations for pediatric radiologic imaging study.

Purpose: The management of geriatric femoral neck fractures, which includes options like hemiarthroplasty (HA), total hip arthroplasty (THA), and fixation, exhibits regional and healthcare setting variations. However, there is a lack of information on global variations in practice patterns and surgical decision factors for this injury., Methods: Survey data were collected from April 2020 to June 2023 via Orthobullets Case Studies, a global clinical case collaboration platform hosted on a prominent orthopedic educational website. Collaboratively developed standardized polls, based on the best available evidence and a comprehensive, peer-reviewed, evidence-based item list, were used to capture surgeons' treatment preferences worldwide. Subsequent analyses explored preferences within subspecialties and practice settings. Multivariable regression analysis identified associations between subspecialty, practice type, the likelihood of choosing THA, and the preferred femoral fixation method., Results: Our study encompassed 2595 respondents from 76 countries. Notably, 51.5% of participants (n = 1328; 51.5%, 95% CI 49.6-53.4%) leaned towards THA and 44.9% for HA, while 3.6% favoured surgical fixation. Respondents affiliated with academic institutions and large non-university-affiliated hospitals were 1.74 times more likely to favour THA, and arthroplasty specialists exhibited a 1.77-fold preference for THA. There was a 19-fold variation for cemented femoral fixation between the United Kingdom (UK) and USA with the UK favouring cemented fixation., Conclusion: Our study reveals a significant shift towards THA preference for managing geriatric femoral neck fractures, influenced by subspecialty and practice settings. We also observed a pronounced predominance of cement fixation in specific geographic locations. These findings highlight the evolving fracture management landscape, emphasizing the need for standardization and comprehensive understanding across diverse healthcare settings., (© 2024. The Author(s), under exclusive licence to Springer-Verlag France SAS, part of Springer Nature.)

Using directed evolution, aminoacyl-tRNA synthetases (aaRSs) have been engineered to incorporate numerous noncanonical amino acids (ncAAs). Until now, the selection of such novel aaRS mutants has relied on the expression of a selectable reporter protein. However, such translation-dependent selections are incompatible with exotic monomers that are suboptimal substrates for the ribosome. A two-step solution is needed to overcome this limitation: (A) engineering an aaRS to charge the exotic monomer, without ribosomal translation; (B) subsequent engineering of the ribosome to accept the resulting acyl-tRNA for translation. Here, we report a platform for aaRS engineering that directly selects tRNA-acylation without ribosomal translation (START). In START, each distinct aaRS mutant is correlated to a cognate tRNA containing a unique sequence barcode. Acylation by an active aaRS mutant protects the corresponding barcode-containing tRNAs from oxidative treatment designed to damage the 3'-terminus of the uncharged tRNAs. Sequencing of these surviving barcode-containing tRNAs is then used to reveal the identity of the aaRS mutants that acylated the correlated tRNA sequences. The efficacy of START was demonstrated by identifying novel mutants of the Methanomethylophilus alvus pyrrolysyl-tRNA synthetase from a naïve library that enables incorporation of ncAAs into proteins in living cells., Competing Interests: The authors declare the following competing financial interest(s): AC and CS are co-inventors on a patent application that incorporates methods outlined in this manuscript. AC is a co-founder and senior advisor at BrickBio, Inc., (© 2024 The Authors. Published by American Chemical Society.)

The development of dendritic cells (DCs), including antigen-presenting conventional DCs (cDCs) and cytokine-producing plasmacytoid DCs (pDCs), is controlled by the growth factor Flt3 ligand (Flt3L) and its receptor Flt3. We genetically dissected Flt3L-driven DC differentiation using CRISPR-Cas9-based screening. Genome-wide screening identified multiple regulators of DC differentiation including subunits of TSC and GATOR1 complexes, which restricted progenitor growth but enabled DC differentiation by inhibiting mTOR signaling. An orthogonal screen identified the transcriptional repressor Trim33 (TIF-1γ) as a regulator of DC differentiation. Conditional targeting in vivo revealed an essential role of Trim33 in the development of all DCs, but not of monocytes or granulocytes. In particular, deletion of Trim33 caused rapid loss of DC progenitors, pDCs, and the cross-presenting cDC1 subset. Trim33-deficient Flt3 + progenitors up-regulated pro-inflammatory and macrophage-specific genes but failed to induce the DC differentiation program. Collectively, these data elucidate mechanisms that control Flt3L-driven differentiation of the entire DC lineage and identify Trim33 as its essential regulator.

Osteoarthritis (OA) is a prevalent joint disease, particularly affecting the knees. This condition is often managed through various treatments, including intra-articular injections such as corticosteroids (CS), hyaluronic acid (HA), and platelet-rich plasma (PRP). PRP has shown promising outcomes in recent studies although it does lack strong endorsement in some clinical guidelines due to inconsistent results and lack of standardized results. This study was conducted to assess patient awareness and the frequency of PRP offered for the treatment of knee OA, compared to CS and HA. In a cross-sectional study, 46 knee OA patients were surveyed regarding their knowledge and experiences of CS, HA, and PRP injections. The questionnaires were administered between September 2022 and February 2023. Additionally, the study evaluated the severity of patients knee OA, using the Western Ontario and McMaster Universities Arthritis Index, and gathered demographic information from the participants. CS injections were offered to 93.5%, and 100% of participants had previously heard of this type of injection. HA injections were offered to 37%, and 65.9% of participants had heard of them. PRP was offered to 2%, and 6.5% had ever heard of it. This study underscores the limited awareness and utilization of PRP among knee OA patients. Patients and physicians need to be more informed of all of the treatment options available for knee OA, especially orthobiologics such as PRP. Future research in larger, diverse populations is needed., Competing Interests: The authors have no funding and conflicts of interest to disclose., (Copyright © 2024 the Author(s). Published by Wolters Kluwer Health, Inc.)

Autoantibodies to chromatin and dsDNA are a hallmark of systemic lupus erythematosus (SLE). In a mouse model of monogenic human SLE caused by DNASE1L3 deficiency, the anti-DNA response is dependent on endosomal nucleic acid-sensing TLRs TLR7 and TLR9. In this study, we report that this response also required TLR2, a surface receptor for microbial products that is primarily expressed on myeloid cells. Cell transfers into lymphopenic DNASE1L3-deficient mice showed that TLR2 was required for anti-DNA Ab production by lymphocytes. TLR2 was detectably expressed on B cells and facilitated the production of IL-6 by B cells activated in the presence of microbial products. Accordingly, treatment with broad-spectrum antibiotics or Ab-mediated blockade of IL-6 delayed the anti-DNA response in DNASE1L3-deficient mice. These studies reveal an unexpected B cell-intrinsic role of TLR2 in systemic autoreactivity to DNA, and they suggest that microbial products may synergize with self-DNA in the activation of autoreactive B cells in SLE., (Copyright © 2023 by The American Association of Immunologists, Inc.)

The complement system is a critical part of our innate immune response, and the terminal products of this cascade, anaphylatoxins C3a and C5a, exert their physiological and pathophysiological responses primarily via two GPCRs, C3aR and C5aR1. However, the molecular mechanism of ligand recognition, activation, and signaling bias of these receptors remains mostly elusive. Here, we present nine cryo-EM structures of C3aR and C5aR1 activated by their natural and synthetic agonists, which reveal distinct binding pocket topologies of complement anaphylatoxins and provide key insights into receptor activation and transducer coupling. We also uncover the structural basis of a naturally occurring mechanism to dampen the inflammatory response of C5a via proteolytic cleavage of the terminal arginine and the G-protein signaling bias elicited by a peptide agonist of C3aR identified here. In summary, our study elucidates the innerworkings of the complement anaphylatoxin receptors and should facilitate structure-guided drug discovery to target these receptors in a spectrum of disorders., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2023 The Author(s). Published by Elsevier Inc. All rights reserved.)

Sulfation widely exists in the eukaryotic proteome. However, understanding the biological functions of sulfation in peptides and proteins has been hampered by the lack of methods to control its spatial or temporal distribution in the proteome. Herein, we report that fluorosulfate can serve as a latent precursor of sulfate in peptides and proteins, which can be efficiently converted to sulfate by hydroxamic acid reagents under physiologically relevant conditions. Photocaging the hydroxamic acid reagents further allowed for the light-controlled activation of functional sulfopeptides. This work provides a valuable tool for probing the functional roles of sulfation in peptides and proteins.

Extracellular DNase DNASE1L3 maintains tolerance to self-DNA in humans and mice, whereas the role of its homolog DNASE1 remains controversial, and the overall function of secreted DNases in immunity is unclear. We report that deletion of murine DNASE1 neither caused autoreactivity in isolation nor exacerbated lupus-like disease in DNASE1L3-deficient mice. However, combined deficiency of DNASE1 and DNASE1L3 rendered mice susceptible to bloodstream infection with Staphylococcus aureus. DNASE1/DNASE1L3 double-deficient mice mounted a normal innate response to S. aureus and did not accumulate neutrophil extracellular traps (NETs). However, their kidneys manifested severe pathology, increased bacterial burden, and biofilm-like bacterial lesions that contained bacterial DNA and excluded neutrophils. Furthermore, systemic administration of recombinant DNASE1 protein during S. aureus infection rescued the mortality of DNase-deficient mice and ameliorated the disease in wild-type mice. Thus, DNASE1 and DNASE1L3 jointly facilitate the control of bacterial infection by digesting extracellular microbial DNA in biofilms, suggesting the original evolutionary function of secreted DNases as antimicrobial agents., (© 2023 Lacey et al.)

The Escherichia coli tyrosyl-tRNA synthetase (EcTyrRS)/tRNA EcTyr pair offers an attractive platform for genetically encoding new noncanonical amino acids (ncAA) in eukaryotes. However, challenges associated with a eukaryotic selection system, which is needed to engineer the platform, have impeded its success in the past. Recently, using a facile E. coli -based selection system, we showed that EcTyrRS could be engineered in a strain where the endogenous tyrosyl pair was substituted with an archaeal counterpart. However, significant cross-reactivity between the UAG-suppressing tRNA CUA EcTyr and the bacterial glutaminyl-tRNA synthetase limited the scope of this strategy, preventing the selection of moderately active EcTyrRS mutants. Here we report an engineered tRNA CUA EcTyr that overcomes this cross-reactivity. Optimized selection systems based on this tRNA enabled the efficient enrichment of both strongly and weakly active ncAA-selective EcTyrRS mutants. We also developed a wide dynamic range (WiDR) antibiotic selection to further enhance the activities of the weaker first-generation EcTyrRS mutants. We demonstrated the utility of our platform by developing several new EcTyrRS mutants that efficiently incorporated useful ncAAs in mammalian cells, including photoaffinity probes, bioconjugation handles, and a nonhydrolyzable mimic of phosphotyrosine., Competing Interests: The authors declare the following competing financial interest(s): A patent application on the EcTyrRS mutants and ncAAs reported here has been submitted. A.C. is a cofounder and senior advisor of BrickBio, Inc., which focuses on applications of the ncAA mutagenesis technology., (© 2022 The Authors. Published by American Chemical Society.)

Antibodies to double-stranded DNA (dsDNA) are prevalent in systemic lupus erythematosus (SLE), particularly in patients with lupus nephritis, yet the nature and regulation of antigenic cell-free DNA (cfDNA) are poorly understood. Null mutations in the secreted DNase DNASE1L3 cause human monogenic SLE with anti-dsDNA autoreactivity. We report that >50% of sporadic SLE patients with nephritis manifested reduced DNASE1L3 activity in circulation, which was associated with neutralizing autoantibodies to DNASE1L3. These patients had normal total plasma cfDNA levels but showed accumulation of cfDNA in circulating microparticles. Microparticle-associated cfDNA contained a higher fraction of longer polynucleosomal cfDNA fragments, which bound autoantibodies with higher affinity than mononucleosomal fragments. Autoantibodies to DNASE1L3-sensitive antigens on microparticles were prevalent in SLE nephritis patients and correlated with the accumulation of cfDNA in microparticles and with disease severity. DNASE1L3-sensitive antigens included DNA-associated proteins such as HMGB1. Our results reveal autoantibody-mediated impairment of DNASE1L3 activity as a common nongenetic mechanism facilitating anti-dsDNA autoreactivity in patients with severe sporadic SLE., Competing Interests: Disclosures: P. Izmirly reported personal fees from GlaxoSmithKline outside the submitted work. J.P. Buyon reported personal fees from Bristol-Myers Squibb LN advisory board, personal fees from GSK panel on LN, personal fees from Amgen, and personal fees from Ventus outside the submitted work. B. Reizis reported being an advisor at Related Sciences. No other disclosures were reported., (© 2021 Hartl et al.)

Detection of microbial nucleic acids by the innate immune system is mediated by numerous intracellular nucleic acids sensors. Upon the detection of nucleic acids these sensors induce the production of inflammatory cytokines, and thus play a crucial role in the activation of anti-microbial immunity. In addition to microbial genetic material, nucleic acid sensors can also recognize self-nucleic acids exposed extracellularly during turn-over of cells, inefficient efferocytosis, or intracellularly upon mislocalization. Safeguard mechanisms have evolved to dispose of such self-nucleic acids to impede the development of autoinflammatory and autoimmune responses. These safeguard mechanisms involve nucleases that are either specific to DNA (DNases) or RNA (RNases) as well as nucleic acid editing enzymes, whose biochemical properties, expression profiles, functions and mechanisms of action will be detailed in this review. Fully elucidating the role of these enzymes in degrading and/or processing of self-nucleic acids to thwart their immunostimulatory potential is of utmost importance to develop novel therapeutic strategies for patients affected by inflammatory and autoimmune diseases., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Santa, Garreau, Serpas, Ferriere, Blanco, Soni and Sisirak.)

Background: Health care technologies are now offering accountability, quality, robustness, and accuracy in disease surveillance and health care delivery programmes. With the advent of mobile hand-held devices, these technologies have become more accessible and adaptable for use by field staff working in remote areas. The Malaria Elimination Demonstration Project started collection of data and conduct of routine operations using paper-based reporting systems. Observing the need for a robust and quality digital mobile application, a comprehensive mobile application tool was developed that allowed the project to conduct disease surveillance, workforce management and supply chain management., Methods: In June 2017, the project conceptualized a comprehensive mobile application tool in the local language (Hindi) for disease surveillance, human resources management, and supply chain management. The tool is also available in English. Solution for Community Health-workers (SOCH) mobile app is an android native application developed using android SDK and web-based tool using MVC.net framework. Construction of the application started in November 2017 and rolled out its pilot in April 2018, followed by pan-district roll out in July 2018. The application uses self-validation tools to ensure high level of data quality and integrity., Results: The software is available in android based hand-held devices and web-screens with built-in data analytical capabilities. Using SOCH, the project has now successfully digitized its routine surveillance, attendance, tour plans, supply chain management components. The project has documented a reduction in 91% indigenous cases in the district, 60% improvement in stock accountability, and 99.6% accuracy in data collected through the mobile application., Conclusion: SOCH is an excellent and user-friendly tool, which can be customized for any public health management programme. The system ensures accountability and data robustness, which is needed for malaria elimination efforts throughout the country. The mobile application can be adapted for English or any other Indian or international language for use for malaria or any other disease surveillance and control programme. Another expansion feature of this mobile application is incorporation of indicators for Indoor Residual Spraying (IRS), Long-Lasting Insecticidal Nets (LLINs), and minor engineering by the residents of community under surveillance. The authors believe that it would be highly desirable and appropriate for an international organization, such as the World Health Organization (WHO), to conduct an independent comparison of all available mobile e-surveillance tools, so that a high-performing and globally suitable system can be selected for use in malaria elimination programmes. The Foundation of Disease Elimination and Controlof India has decided to make the SOCH mobile application available to anyone who would like to use it for disease surveillance and health care programmes.

IL-10 producing B cells (B10 cells) play an important immunoregulatory role in various autoimmune and infection conditions. However, the factors that regulate their development and maintenance are incompletely understood. Recently, we and others have established a requirement for TLR7 in promoting autoimmune antibody forming cell (AFC) and germinal center (GC) responses. Here we report an important additional role of TLR7 in the negative regulation of B10 cell development. TLR7 overexpression or overstimulation promoted the reduction of B10 cells whereas TLR7 deficiency rescued these cells in both non-autoimmune and autoimmune-prone mice. TLR7 expression was further inversely correlated with B cell-dependent IL-10 production and its inhibition of CD4 T cell proliferation and IFNγ production in an in vitro B cell and T cell co-culture system. Further, B10 cells displayed elevated TLR7, IFNγR, and STAT1 expression compared to non-B10 cells. Interestingly, deficiency of IFNγR in TLR7 overexpressing lupus-prone mice rescued B10 cells from TLR7-mediated reduction. Finally, B cell intrinsic deletion of IFNγR was sufficient to restore B10 cells in the spleens of TLR7-promoted autoimmune mouse model. In conclusion, our findings demonstrate a novel role for the IFNγR-STAT1 pathway in TLR7-mediated negative regulation of B10 cell development., (Copyright © 2020 Chodisetti, Fike, Domeier, Choi, Soni and Rahman.)

Class-switched antibodies to double-stranded DNA (dsDNA) are prevalent and pathogenic in systemic lupus erythematosus (SLE), yet mechanisms of their development remain poorly understood. Humans and mice lacking secreted DNase DNASE1L3 develop rapid anti-dsDNA antibody responses and SLE-like disease. We report that anti-DNA responses in Dnase1l3 -/- mice require CD40L-mediated T cell help, but proceed independently of germinal center formation via short-lived antibody-forming cells (AFCs) localized to extrafollicular regions. Type I interferon (IFN-I) signaling and IFN-I-producing plasmacytoid dendritic cells (pDCs) facilitate the differentiation of DNA-reactive AFCs in vivo and in vitro and are required for downstream manifestations of autoimmunity. Moreover, the endosomal DNA sensor TLR9 promotes anti-dsDNA responses and SLE-like disease in Dnase1l3 -/- mice redundantly with another nucleic acid-sensing receptor, TLR7. These results establish extrafollicular B cell differentiation into short-lived AFCs as a key mechanism of anti-DNA autoreactivity and reveal a major contribution of pDCs, endosomal Toll-like receptors (TLRs), and IFN-I to this pathway., Competing Interests: Declaration of Interests The authors declare no competing interests., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Intestinal mononuclear phagocytes (MPs) are composed of heterogeneous dendritic cell (DC) and macrophage subsets necessary for the initiation of immune response and control of inflammation. Although MPs in the normal intestine have been extensively studied, the heterogeneity and function of inflammatory MPs remain poorly defined. We performed phenotypical, transcriptional, and functional analyses of inflammatory MPs in infectious Salmonella colitis and identified CX3CR1 + MPs as the most prevalent inflammatory cell type. CX3CR1 + MPs were further divided into three distinct populations, namely, Nos2 + CX3CR1 lo , Ccr7 + CX3CR1 int (lymph migratory), and Cxcl13 + CX3CR1 hi (mucosa resident), all of which were transcriptionally aligned with macrophages and derived from monocytes. In follow-up experiments in vivo, intestinal CX3CR1 + macrophages were superior to conventional DC1 (cDC1) and cDC2 in inducing Salmonella -specific mucosal IgA. We next examined spatial organization of the immune response induced by CX3CR1 + macrophage subsets and identified mucosa-resident Cxcl13 + CX3CR1 hi macrophages as the antigen-presenting cells responsible for recruitment and activation of CD4 + T and B cells to the sites of Salmonella invasion, followed by tertiary lymphoid structure formation and the local pathogen-specific IgA response. Using mice we developed with a floxed Ccr7 allele, we showed that this local IgA response developed independently of migration of the Ccr7 + CX3CR1 int population to the mesenteric lymph nodes and contributed to the total mucosal IgA response to infection. The differential activity of intestinal macrophage subsets in promoting mucosal IgA responses should be considered in the development of vaccines to prevent Salmonella infection and in the design of anti-inflammatory therapies aimed at modulating macrophage function in inflammatory bowel disease., (Copyright © 2020 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works.)