"You should brush your teeth better": a randomized controlled trial comparing best-possible versus as-usual toothbrushing.

BACKGROUND: Most people's tooth brushing performance is deficient, even when they are encouraged to brush to the best of their abilities. The aim of the present study was to explore the nature of this deficit by comparing best-possible vs. as-usual brushing. METHODS: University students (N = 111) were randomly assigned to receive one of two instructions: "brush your teeth as usual" (AU-instruction) or "brush your teeth to the best of your abilities" (BP-instruction). Video analyses assessed brushing performance. The marginal plaque index (MPI) assessed after brushing was used as an indicator of brushing effectiveness. A questionnaire assessed subjectively perceived oral cleanliness (SPOC). RESULTS: Participants in the BP group brushed their teeth longer (p = 0.008, d = 0.57) and used interdental devices more often (p < 0.001). No group differences emerged in the distribution of brushing time among surfaces, the percentage of brushing techniques used beyond horizontal scrubbing, or the appropriate use of interdental devices (all p > 0.16, all d < 0.30). Plaque persisted at the majority of the sections of the gingival margins, and the groups did not differ in this respect (p = 0.15; d = 0.22). SPOC values in the BP group were higher than those in the AU group (p = 0.006; d = 0.54). Both groups overestimated their actual oral cleanliness by approximately twofold. CONCLUSIONS: Compared to their usual brushing effort, study participants increased their effort when asked to brush their teeth in the best possible manner. However, that increase in effort was ineffective in terms of oral cleanliness. The results indicate that people's concept of optimized brushing refers to quantitative aspects (e.g., longer duration, more interdental hygiene) rather than qualitative aspects (e.g., considering inner surfaces and gingival margins, appropriate use of dental floss). TRIAL REGISTRATION: The study was registered in the appropriate national register ( www.drks.de ; ID: DRKS00017812; date of registration: 27/08/2019 - retrospectively registered).

Good role models? Tooth brushing capabilities of parents: a video observation study.

BACKGROUND: Research in adolescents reveals that they are not capable to remove dental plaque effectively. Inconsistent application of brushing techniques and neglect of certain areas while brushing are very common. As parents play a major role in the oral health education, the present study aimed to examine and describe the tooth brushing performance of the parents of adolescents. METHODS: Parents of adolescents (N = 66) were asked to perform oral hygiene to the best of their capabilities in front of a video camera and dental plaque was recorded before and afterwards. Papillary bleeding was also assessed. RESULTS: The tooth contact time (i.e. net brushing duration) averaged 155 s ± 58 s. Gingival margins showed persistent plaque at 68% ± 14% of the sections assessed. Papillary bleeding was found at 52% ± 18% of the papillae. Parents brushed inner surfaces lesser than the outer surfaces (41 s ± 24 s vs. 73 s ± 33 s) and 29% of the parents completely missed the inner surfaces of at least one sextant. On the outer surfaces parents predominantly applied circular movements (66% ± 25% of brushing time). Vertical brushing prevailed on the inner surfaces (52% ± 30%). However, horizontal scrubbing was also very common (46% ± 31%). CONCLUSIONS: Parents' tooth brushing performance was neither effective in terms of plaque removal nor did they fully comply with tooth brushing recommendations such as considering all inner surfaces when brushing or application of other than horizontal movements to lateral surfaces. Regarding oral hygiene, parents should not only be a good role model in terms of timing, frequency and duration of tooth brushing but should also be able to demonstrate how to brush teeth completely and effectively. The current research indicates that they might lack the latter skill.

Tooth brushing performance in adolescents as compared to the best-practice demonstrated in group prophylaxis programs: an observational study.

BACKGROUND: Research indicates that adolescents may have difficulties to adopt the tooth brushing recommendations delivered in prophylaxis programs. However, it is not clear whether these difficulties are seen amongst the entire age range of adolescence (10-19 years) or only occur at certain developmental stages of the adolescence. The present study analyzes the tooth brushing performance of adolescents and compares it to the best-practice of tooth brushing demonstrated during prophylaxis programs. METHODS: A random sample of N = 66 adolescents, comprising 10-year-olds (n = 42) and 15-year-olds (n = 24), were asked to perform oral hygiene to the best of their abilities in front of a tablet camera. Videos were analyzed for tooth brushing duration, location, and brushing movements, and the difference between the actual and expected behaviour was tested for consistency using repeated measures ANOVAs and Student's t-tests. For the direct comparison across different age groups, already available data from 12- and 18-year-olds were reanalysed. RESULTS: The average brushing time (mean ± SD) of the 10-year-olds and 15-year-olds was 195.8 s (74.6 s) and 196.1 s (75.8 s), respectively. Regardless of age, the adolescents distributed their brushing time unevenly across the inner, outer and occlusal surfaces. The inner surfaces in particular were neglected to a considerable extent, as no age group spent more than 15.8% of the total brushing time on them. Furthermore, all age groups showed a high proportion of horizontal movements on the inner and outer surfaces, regardless of the movements instructed for the respective surfaces. CONCLUSION: Even if adolescents brush to the best of their abilities, they neglect or skip one or many of the tooth surfaces. The reasons for the lack of compliance to tooth brushing instructions are discussed in light of the methods used in prophylaxis programs and the influence of parents.

Vaccination status and COVID-19 related mortality: A hospital based cross sectional study.

BACKGROUND: Efficacy of vaccines studied in clinical trial settings are likely to be different from their effectiveness in a real-world scenario. Indian Armed Forces launched its vaccine drive against COVID-19 on 16 Jan 2021. This study evaluated the effect of vaccination on mortality amongst hospitalized COVID patients. METHODS: A cross sectional study was done on all admitted moderate to severe COVID-19 patients at a designated COVID hospital in New Delhi. The primary outcome assessed the association of being fully vaccinated with mortality. Unadjusted odds ratios (OR) (with 95% CI) was performed for each predictor. Logistic regression was used for multivariable analysis and adjusted odds ratios obtained. RESULTS: The 1168 patients included in the study had a male preponderance with a mean age of 54.6 (± 17.51) years. A total of 266 (23%) patients were partially vaccinated with COVISHIELD® and 184 (16%) were fully vaccinated. Overall, 518 (44.3%) patients had comorbidities and 332 (28.4%) died. Among those fully vaccinated, there was 12.5% (23/184) mortality while it was 31.45 % (309/984) among the unvaccinated (OR 0.3, 95% CI 0.2 to 0.5, p<0.0001). In a logistic regression model, complete vaccination status and younger age were found to be associated with survival. CONCLUSIONS: Vaccination with two doses of COVISHIELD® was associated with lower odds of mortality among hospitalized patients with moderate to severe COVID.

Systematic review of predictive mathematical models of COVID-19 epidemic.

BACKGROUND: Various mathematical models were published to predict the epidemiological consequences of the COVID-19 pandemic. This systematic review has studied the initial epidemiological models. METHODS: Articles published from January to June 2020 were extracted from databases using search strings and those peer-reviewed with full text in English were included in the study. They were analysed as to whether they made definite predictions in terms of time and numbers, or contained only mathematical assumptions and open-ended predictions. Factors such as early vs. late prediction models, long-term vs. curve-fitting models and comparisons based on modelling techniques were analysed in detail. RESULTS: Among 56,922 hits in 05 databases, screening yielded 434 abstracts, of which 72 articles were included. Predictive models comprised over 70% (51/72) of the articles, with susceptible, exposed, infectious and recovered (SEIR) being the commonest type (mean duration of prediction being 3 months). Common predictions were regarding cumulative cases (44/72, 61.1%), time to reach total numbers (41/72, 56.9%), peak numbers (22/72, 30.5%), time to peak (24/72, 33.3%), hospital utilisation (7/72, 9.7%) and effect of lockdown and NPIs (50/72, 69.4%). The commonest countries for which models were predicted were China followed by USA, South Korea, Japan and India. Models were published by various professionals including Engineers (12.5%), Mathematicians (9.7%), Epidemiologists (11.1%) and Physicians (9.7%) with a third (32.9%) being the result of collaborative efforts between two or more professions. CONCLUSION: There was a wide diversity in the type of models, duration of prediction and the variable that they predicted, with SEIR model being the commonest type.

COVISHIELD (AZD1222) VaccINe effectiveness among healthcare and frontline Workers of INdian Armed Forces: Interim results of VIN-WIN cohort study.

BACKGROUND: On 16 Jan 2021, India launched its immunization program against COVID-19. Among the first recipients were 1.59 million Health Care Workers (HCWs) and Frontline Workers (FLWs) of the Indian Armed Forces, who were administered COVISHIELD (Astra Zeneca). We present an interim analysis of vaccine effectiveness (VE) estimates till 30 May 2021. METHODS: The VIN-WIN cohort study was carried out on anonymized data of HCWs and FLWs of Indian Armed Forces. The existing surveillance system, enhanced for COVID-19 monitoring, was sourced for data. The cohort transitioned from Unvaccinated (UV) to Partially Vaccinated (PV) to Fully Vaccinated (FV), serving as its own internal comparison. Outcomes studied in the three groups were breakthrough infections and COVID related deaths. Incidence Rate Ratio (IRR) was used to compare outcomes among the three groups to estimate VE. RESULTS: Data of 1,595,630 individuals (mean age 27.6 years; 99% male) over 135 days was analysed. Till 30 May 21, 95.4% and 82.2% were partially and fully vaccinated. The UV, PV and FV compartments comprised 106.6, 46.7 and 58.7 million person-days respectively. The number of breakthrough cases in the UV, PV and FV groups were 10061, 1159 and 2512; while the deaths were 37, 16 and 7 respectively. Corrected VE was 91.8-94.9% against infections. CONCLUSION: Interim results of the VIN-WIN cohort study of 1.59 million HCWs and FLWs of Indian Armed Forces showed a ∼93% reduction in COVID-19 breakthrough infections with COVISHIELD vaccination.

Healthcare impact of COVID-19 epidemic in India: A stochastic mathematical model.

BACKGROUND: In India, the SARS-CoV-2 COVID-19 epidemic has grown to 1251 cases and 32 deaths as on 30 Mar 2020. The healthcare impact of the epidemic in India was studied using a stochastic mathematical model. METHODS: A compartmental SEIR model was developed, in which the flow of individuals through compartments is modeled using a set of differential equations. Different scenarios were modeled with 1000 runs of Monte Carlo simulation each using MATLAB. Hospitalization, intensive care unit (ICU) requirements, and deaths were modeled on SimVoi software. The impact of nonpharmacological interventions (NPIs) including social distancing and lockdown on checking the epidemic was estimated. RESULTS: Uninterrupted epidemic in India would have resulted in more than 364 million cases and 1.56 million deaths with peak by mid-July. As per the model, at current growth rate of 1.15, India is likely to reach approximately 3 million cases by 25 May, implying 125,455 (±18,034) hospitalizations, 26,130 (±3298) ICU admissions, and 13,447 (±1819) deaths. This would overwhelm India's healthcare system. The model shows that with immediate institution of NPIs, the epidemic might still be checked by mid-April 2020. It would then result in 241,974 (±33,735) total infections, 10,214 (±1649) hospitalizations, 2121 (±334) ICU admissions, and 1081 (±169) deaths. CONCLUSION: At the current growth rate of epidemic, India's healthcare resources will be overwhelmed by the end of May. With the immediate institution of NPIs, total cases, hospitalizations, ICU requirements, and deaths can be reduced by almost 90%.

Coronavirus disease 2019 in India: Post-lockdown scenarios and provisioning for health care.

BACKGROUND: With the rise of Coronavirus disease 2019 (COVID-19) cases in India, lockdown was imposed from March 25, 2020. We studied post-lockdown scenarios and evaluated health-care constraints. Our aim was to identify the scenarios in which health-care availability would not be overwhelmed. METHODS: A modified compartmental SEIR stochastic model was used to calculate peak cases at various levels of effectiveness of prevention of transmission. Health-care constraints were evaluated using a Delphi study. We developed "q-metric" to evaluate the epidemic. Key constraints were matched against scenarios generated, and a monitoring mechanism was devised. RESULTS: Continuing lockdown ("q-metric" of >50) until mid-August was theoretically the most effective solution to end the epidemic. Lockdown might however be lifted earlier owing to various compulsions. The key constraints were identified as trained manpower and ventilators. It was estimated that shortfall of specialists to operate ventilators for COVID-19 intensive care units was approximately 40,000. This requires re-purposing of other specialists and short-term training to meet the surge. The shortage of ventilators is around 40,000-50,000. Procuring beyond those numbers would be infructuous owing to limits of training manpower. After lifting lockdown, the aim should be to contain the epidemic within the availability of key constraints. Our model suggests that this can be achieved by community containment and other non-pharmacological interventions at a "q-metric" of 19. An algorithm using "q-metric" was developed to monitor the epidemic. CONCLUSION: Various post-lockdown scenarios were simulated. Trained manpower and ventilators were identified as key health-care constraints. Partial community containment measures will require to be continued after the current lockdown is lifted.

Predictors of first line antiretroviral therapy failure and burden of second line antiretroviral therapy.

BACKGROUND: As HIV steps into the third decade, there are more number of patients living on lifelong (antiretroviral therapy) ART and facing the threat of drug resistance with subsequent treatment failure. The aim of this study was to determine predictors of first-line ART failure with the objectives to estimate the burden of 2nd line ART. METHODS: A retrospective 5-year cohort of HIV patients who were initiated on first line ART in 2008-09 was studied. Patients were followed from the time of ART initiation. Kaplan-Meier methods and Cox proportional hazards regression models were used to estimate probabilities and predictors of first line ART failure. RESULTS: Of the total of 195 patients initiated on first line ART, 15 patients were switched to second line ART yielding 7.69% failure rate. During the 7178 person-years of follow-up, the incidence of first line ART failure was 2.09 per 1000 person-years. The Kaplan-Meier survival analysis gave a mean survival time of 55.6 months. BMI, CD4 count at ART initiation and presence of opportunistic infections were significant predictors of first line ART failure. The burden of second line ART patients by the end of 5 years of first line ART is expected to be 151 patients. CONCLUSION: Though the first line ART failure is quite low in this study, we still need to be vigilant for lower BMI, low baseline CD4 count and occurrence of opportunistic infections to efficiently manage failures on first line ART.

Synovial fluid proteome in rheumatoid arthritis.

BACKGROUND: Rheumatoid arthritis (RA) is a chronic autoinflammatory disorder that affects small joints. Despite intense efforts, there are currently no definitive markers for early diagnosis of RA and for monitoring the progression of this disease, though some of the markers like anti CCP antibodies and anti vimentin antibodies are promising. We sought to catalogue the proteins present in the synovial fluid of patients with RA. It was done with the aim of identifying newer biomarkers, if any, that might prove promising in future. METHODS: To enrich the low abundance proteins, we undertook two approaches-multiple affinity removal system (MARS14) to deplete some of the most abundant proteins and lectin affinity chromatography for enrichment of glycoproteins. The peptides were analyzed by LC-MS/MS on a high resolution Fourier transform mass spectrometer. RESULTS: This effort was the first total profiling of the synovial fluid proteome in RA that led to identification of 956 proteins. From the list, we identified a number of functionally significant proteins including vascular cell adhesion molecule-1, S100 proteins, AXL receptor protein tyrosine kinase, macrophage colony stimulating factor (M-CSF), programmed cell death ligand 2 (PDCD1LG2), TNF receptor 2, (TNFRSF1B) and many novel proteins including hyaluronan-binding protein 2, semaphorin 4A (SEMA4D) and osteoclast stimulating factor 1. Overall, our findings illustrate the complex and dynamic nature of RA in which multiple pathways seems to be participating actively. CONCLUSIONS: The use of high resolution mass spectrometry thus, enabled identification of proteins which might be critical to the progression of RA.

Plasma Proteome Database as a resource for proteomics research: 2014 update.

Plasma Proteome Database (PPD; http://www.plasmaproteomedatabase.org/) was initially described in the year 2005 as a part of Human Proteome Organization's (HUPO's) pilot initiative on Human Plasma Proteome Project. Since then, improvements in proteomic technologies and increased throughput have led to identification of a large number of novel plasma proteins. To keep up with this increase in data, we have significantly enriched the proteomic information in PPD. This database currently contains information on 10,546 proteins detected in serum/plasma of which 3784 have been reported in two or more studies. The latest version of the database also incorporates mass spectrometry-derived data including experimentally verified proteotypic peptides used for multiple reaction monitoring assays. Other novel features include published plasma/serum concentrations for 1278 proteins along with a separate category of plasma-derived extracellular vesicle proteins. As plasma proteins have become a major thrust in the field of biomarkers, we have enabled a batch-based query designated Plasma Proteome Explorer, which will permit the users in screening a list of proteins or peptides against known plasma proteins to assess novelty of their data set. We believe that PPD will facilitate both clinical and basic research by serving as a comprehensive reference of plasma proteins in humans and accelerate biomarker discovery and translation efforts.

Sociodemographic characteristics and aggression quotient among children in conflict with the law in India: A case-control study.

BACKGROUND: The incidence of juvenile delinquency has increased in the past decade in India and juvenile crimes are increasingly being reported. This has been attributed to many biopsychosocial factors. It is essential to understand these issues in the context of India to determine the response of acts of children in conflict with the law. We aimed to assess the sociodemographic characteristics and the aggression quotient of children in conflict with the law (juvenile delinquents) in observation homes across India and compare them with those who were not. METHODS: We did a case-control study in five juvenile homes in the cities of Hyderabad, Lucknow and Pune. Ninety inmates (74 boys, 16 girls) were included in the study. Sociodemographic characteristics and aggression quotient of children in conflict with the law were evaluated using two separate questionnaires, i.e. a sociodemographic questionnaire and the modified Buss and Perry aggression questionnaire. These were compared with a control group of similar age, sex and income status. RESULTS: All the children in conflict with the law surveyed belonged to the lower socioeconomic strata, had a significantly higher chance (p<0.05) of coming from broken homes, have addictions, jailed family members and suffered physical and sexual abuse than controls. They also scored higher on all domains of the aggression questionnaire than controls. CONCLUSION: A collection of sociodemographic attributes such as broken homes, addictions and abuse seem to have an important association with juvenile delinquency. Children in conflict with the law are also more likely to be associated with a higher aggression quotient as compared to children who were not.

Differential proteomic analysis of synovial fluid from rheumatoid arthritis and osteoarthritis patients.

BACKGROUND: Rheumatoid arthritis and osteoarthritis are two common musculoskeletal disorders that affect the joints. Despite high prevalence rates, etiological factors involved in these disorders remain largely unknown. Dissecting the molecular aspects of these disorders will significantly contribute to improving their diagnosis and clinical management. In order to identify proteins that are differentially expressed between these two conditions, a quantitative proteomic profiling of synovial fluid obtained from rheumatoid arthritis and osteoarthritis patients was carried out by using iTRAQ labeling followed by high resolution mass spectrometry analysis. RESULTS: We have identified 575 proteins out of which 135 proteins were found to be differentially expressed by ≥3-fold in the synovial fluid of rheumatoid arthritis and osteoarthritis patients. Proteins not previously reported to be associated with rheumatoid arthritis including, coronin-1A (CORO1A), fibrinogen like-2 (FGL2), and macrophage capping protein (CAPG) were found to be upregulated in rheumatoid arthritis. Proteins such as CD5 molecule-like protein (CD5L), soluble scavenger receptor cysteine-rich domain-containing protein (SSC5D), and TTK protein kinase (TTK) were found to be upregulated in the synovial fluid of osteoarthritis patients. We confirmed the upregulation of CAPG in rheumatoid arthritis synovial fluid by multiple reaction monitoring assay as well as by Western blot. Pathway analysis of differentially expressed proteins revealed a significant enrichment of genes involved in glycolytic pathway in rheumatoid arthritis. CONCLUSIONS: We report here the largest identification of proteins from the synovial fluid of rheumatoid arthritis and osteoarthritis patients using a quantitative proteomics approach. The novel proteins identified from our study needs to be explored further for their role in the disease pathogenesis of rheumatoid arthritis and osteoarthritis.Sartaj Ahmad and Raja Sekhar Nirujogi contributed equally to this article.

Proteomic analysis of human osteoarthritis synovial fluid.

BACKGROUND: Osteoarthritis is a chronic musculoskeletal disorder characterized mainly by progressive degradation of the hyaline cartilage. Patients with osteoarthritis often postpone seeking medical help, which results in the diagnosis being made at an advanced stage of cartilage destruction. Sustained efforts are needed to identify specific markers that might help in early diagnosis, monitoring disease progression and in improving therapeutic outcomes. We employed a multipronged proteomic approach, which included multiple fractionation strategies followed by high resolution mass spectrometry analysis to explore the proteome of synovial fluid obtained from osteoarthritis patients. In addition to the total proteome, we also enriched glycoproteins from synovial fluid using lectin affinity chromatography. RESULTS: We identified 677 proteins from synovial fluid of patients with osteoarthritis of which 545 proteins have not been previously reported. These novel proteins included ADAM-like decysin 1 (ADAMDEC1), alanyl (membrane) aminopeptidase (ANPEP), CD84, fibulin 1 (FBLN1), matrix remodelling associated 5 (MXRA5), secreted phosphoprotein 2 (SPP2) and spondin 2 (SPON2). We identified 300 proteins using lectin affinity chromatography, including the glycoproteins afamin (AFM), attractin (ATRN), fibrillin 1 (FBN1), transferrin (TF), tissue inhibitor of metalloproteinase 1 (TIMP1) and vasorin (VSN). Gene ontology analysis confirmed that a majority of the identified proteins were extracellular and are mostly involved in cell communication and signaling. We also confirmed the expression of ANPEP, dickkopf WNT signaling pathway inhibitor 3 (DKK3) and osteoglycin (OGN) by multiple reaction monitoring (MRM) analysis of osteoarthritis synovial fluid samples. CONCLUSIONS: We present an in-depth analysis of the synovial fluid proteome from patients with osteoarthritis. We believe that the catalog of proteins generated in this study will further enhance our knowledge regarding the pathophysiology of osteoarthritis and should assist in identifying better biomarkers for early diagnosis.

A multilectin affinity approach for comparative glycoprotein profiling of rheumatoid arthritis and spondyloarthropathy.

BACKGROUND: Arthritis refers to inflammation of joints and includes common disorders such as rheumatoid arthritis (RA) and spondyloarthropathies (SpAs). These diseases differ mainly in terms of their clinical manifestations and the underlying pathogenesis. Glycoproteins in synovial fluid might reflect the disease activity status in the joints affected by arthritis; yet they have not been systematically studied previously. Although markers have been described for assisting in the diagnosis of RA, there are currently no known biomarkers for SpA. MATERIALS AND METHODS: We sought to determine the relative abundance of glycoproteins in RA and SpA by lectin affinity chromatography coupled to iTRAQ labeling and LC-MS/MS analysis. We also used ELISA to validate the overexpression of VCAM-1, one of the candidate proteins identified in this study, in synovial fluid from RA patients. RESULTS AND DISCUSSION: We identified proteins that were previously reported to be overexpressed in RA including metalloproteinase inhibitor 1 (TIMP1), myeloperoxidase (MPO) and several S100 proteins. In addition, we discovered several novel candidates that were overexpressed in SpA including Apolipoproteins C-II and C-III and the SUN domain-containing protein 3 (SUN3). Novel molecules found overexpressed in RA included extracellular matrix protein 1 (ECM1) and lumican (LUM). We validated one of the candidate biomarkers, vascular cell adhesion molecule 1 (VCAM1), in 20 RA and SpA samples using ELISA and confirmed its overexpression in RA (p-value <0.01). Our quantitative glycoproteomic approach to study arthritic disorders should open up new avenues for additional proteomics-based discovery studies in rheumatological disorders.

Role of interleukin 10 transcriptional regulation in inflammation and autoimmune disease.

Interleukin 10 (IL-10) is a cytokine with potent anti-inflammatory properties that plays a central role in limiting host immune response to pathogens, thereby preventing damage to the host and maintaining normal tissue homeostasis. Dysregulation of IL-10 is associated with enhanced immunopathology in response to infection as well as increased risk for development of many autoimmune diseases. Thus a fundamental understanding of IL-10 gene expression is critical for our comprehension of disease progression and resolution of host inflammatory response. In this review, we discuss modes of regulation of IL-10 gene expression in immune effector cell types, including signal transduction, epigenetics, promoter architecture, and post-transcriptional regulation, and how aberrant regulation contributes to immunopathology and disease progression.

All that seems sepsis is not sepsis.

Catastrophic antiphospholipid antibody syndrome (CAPS) resembles severe sepsis in its acute presentation, with features of systemic inflammatory response syndrome (SIRS) leading to multiple organ dysfunction. Infections are the best known triggers of CAPS. This emphasizes the need for early diagnosis and aggressive treatment as the mortality is as high as 50%. We present a 42-year-old woman who developed SIRS postoperatively and was eventually diagnosed as CAPS.

Lipopolysaccharide-mediated IL-10 transcriptional regulation requires sequential induction of type I IFNs and IL-27 in macrophages.

IL-10 is a potent anti-inflammatory molecule that regulates excessive production of inflammatory cytokines during an infection or tissue damage. Dysregulation of IL-10 is associated with a number of autoimmune diseases, and so, understanding the mechanisms by which IL-10 gene expression is regulated remains an important area of study. Macrophages represent a major source of IL-10, which is generated in response to TLR signaling as a feedback mechanism to curtail inflammatory response. In this study, we identify a signaling pathway in murine bone marrow-derived macrophages in which activation of TLR4 by LPS induces the expression of IL-10 through the sequential induction of type I IFNs followed by induction and signaling through IL-27. We demonstrate that IL-27 signaling is required for robust IL-10 induction by LPS and type I IFNs. IL-27 leads directly to transcription of IL-10 through the activation of two required transcription factors, STAT1 and STAT3, which are recruited to the IL-10 promoter. Finally, through systematic functional promoter-reporter analysis, we identify three cis elements within the proximal IL-10 promoter that play an important role in regulating transcription of IL-10 in response to IL-27.