Tim4 enables large peritoneal macrophages to cross-present tumor antigens at early stages of tumorigenesis.

Receptors controlling the cross-presentation of tumor antigens by macrophage subsets in cancer tissues are poorly explored. Here, we show that TIM4+ large peritoneal macrophages efficiently capture and cross-present tumor-associated antigens at early stages of peritoneal infiltration by ovarian cancer cells. The phosphatidylserine (PS) receptor TIM4 promotes maximal uptake of dead cells or PS-coated artificial targets and triggers inflammatory and metabolic gene programs in combination with cytoskeletal remodeling and upregulation of transcriptional signatures related to antigen processing. At the cellular level, TIM4-mediated engulfment induces nucleation of F-actin around nascent phagosomes, delaying the recruitment of vacuolar ATPase, acidification, and cargo degradation. In vivo, TIM4 deletion blunts induction of early anti-tumoral effector CD8 T cells and accelerates the progression of ovarian tumors. We conclude that TIM4-mediated uptake drives the formation of specialized phagosomes that prolong the integrity of ingested antigens and facilitate cross-presentation, contributing to immune surveillance of the peritoneum.

Dendritic cell-targeted therapy expands CD8 T cell responses to bona-fide neoantigens in lung tumors.

Cross-presentation by type 1 DCs (cDC1) is critical to induce and sustain antitumoral CD8 T cell responses to model antigens, in various tumor settings. However, the impact of cross-presenting cDC1 and the potential of DC-based therapies in tumors carrying varied levels of bona-fide neoantigens (neoAgs) remain unclear. Here we develop a hypermutated model of non-small cell lung cancer in female mice, encoding genuine MHC-I neoepitopes to study neoAgs-specific CD8 T cell responses in spontaneous settings and upon Flt3L + αCD40 (DC-therapy). We find that cDC1 are required to generate broad CD8 responses against a range of diverse neoAgs. DC-therapy promotes immunogenicity of weaker neoAgs and strongly inhibits the growth of high tumor-mutational burden (TMB) tumors. In contrast, low TMB tumors respond poorly to DC-therapy, generating mild CD8 T cell responses that are not sufficient to block progression. scRNA transcriptional analysis, immune profiling and functional assays unveil the changes induced by DC-therapy in lung tissues, which comprise accumulation of cDC1 with increased immunostimulatory properties and less exhausted effector CD8 T cells. We conclude that boosting cDC1 activity is critical to broaden the diversity of anti-tumoral CD8 T cell responses and to leverage neoAgs content for therapeutic advantage.

Residency Application Advising: Medical Student & Faculty Advisor Perceived Barriers & Expectations.

INTRODUCTION: Given the rapidly changing landscape of residency applications, many medical students struggle to identify guidance from faculty advisors. Additionally, faculty advisors may find it difficult to maintain up-to-date knowledge on changes such as the new supplemental application. These gaps could potentially lead to inequitable advising. The objective of this study was to identify both students' and faculty's perceived barriers and expectations for residency application advising. METHODS: Anonymous surveys were administered to both fourth-year medical students and faculty advisors at a single institution within 2 mo of the residency application deadline. Survey questions assessed student and faculty barriers to establishing the advisor-advisee relationships, as well as expectations of the advisor role. Surveys were analyzed using descriptive statistics. RESULTS: We identified that the majority of students (57%) did not have a faculty advisor within weeks of the application deadline, and an equal amount felt that finding an advisor was either somewhat difficult or extremely difficult. Of all the students, 60% felt their biggest barrier was not knowing how to find an advisor. Though faculty felt equipped to advise students, 75% of faculty in the participating specialties had advising concerns regarding the supplemental application or were unaware of the changes. CONCLUSIONS: We identified gaps in the residency application advising process from both student and faculty perspectives. Future work involves increasing awareness of the resources and opportunities available to students to improve advising relationships. Standardized training tools and resources for faculty will result in more consistent and reliable faculty advising.

Free fatty acid receptor 4 responds to endogenous fatty acids to protect the heart from pressure overload.

AIMS: Free fatty acid receptor 4 (Ffar4) is a G-protein-coupled receptor for endogenous medium-/long-chain fatty acids that attenuates metabolic disease and inflammation. However, the function of Ffar4 in the heart is unclear. Given its putative beneficial role, we hypothesized that Ffar4 would protect the heart from pathologic stress. METHODS AND RESULTS: In mice lacking Ffar4 (Ffar4KO), we found that Ffar4 is required for an adaptive response to pressure overload induced by transverse aortic constriction (TAC), identifying a novel cardioprotective function for Ffar4. Following TAC, remodelling was worsened in Ffar4KO hearts, with greater hypertrophy and contractile dysfunction. Transcriptome analysis 3-day post-TAC identified transcriptional deficits in genes associated with cytoplasmic phospholipase A2α signalling and oxylipin synthesis and the reduction of oxidative stress in Ffar4KO myocytes. In cultured adult cardiac myocytes, Ffar4 induced the production of the eicosapentaenoic acid (EPA)-derived, pro-resolving oxylipin 18-hydroxyeicosapentaenoic acid (18-HEPE). Furthermore, the activation of Ffar4 attenuated cardiac myocyte death from oxidative stress, while 18-HEPE rescued Ffar4KO myocytes. Systemically, Ffar4 maintained pro-resolving oxylipins and attenuated autoxidation basally, and increased pro-inflammatory and pro-resolving oxylipins, including 18-HEPE, in high-density lipoproteins post-TAC. In humans, Ffar4 expression decreased in heart failure, while the signalling-deficient Ffar4 R270H polymorphism correlated with eccentric remodelling in a large clinical cohort paralleling changes observed in Ffar4KO mice post-TAC. CONCLUSION: Our data indicate that Ffar4 in cardiac myocytes responds to endogenous fatty acids, reducing oxidative injury, and protecting the heart from pathologic stress, with significant translational implications for targeting Ffar4 in cardiovascular disease.

Linguistic history and language diversity in India: Views and counterviews.

This paper addresses the theme of the seminar from the perspective of historical linguistics. It introduces the construct of 'language family' and then proceeds to a discussion of contact and the dynamics of linguistic exchange among the main language families of India over several millennia. Some prevalent hypotheses to explain the creation of India as a linguistic area are presented. The 'substratum view' is critically assessed. Evidence from historical linguistics in support of two dominant hypotheses - 'the Aryan migration view''and 'the out-of-India hypothesis' - is presented and briefly assessed. In conclusion, it is observed that the current understanding in historical linguistics favours the Aryan migration view though the 'substratum view' is questionable.

Mouse Models of Pain in Sickle Cell Disease.

Sickle cell disease (SCD) is a genetic blood disorder that impacts millions of individuals worldwide. SCD is characterized by debilitating pain that can begin during infancy and may continue to increase throughout life. This pain can be both acute and chronic. A characteristic feature specific to acute pain in SCD occurs during vaso-occlusive crisis (VOC) due to the blockade of capillaries with sickle red blood cells. The acute pain of VOC is intense, unpredictable, and requires hospitalization. Chronic pain occurs in a significant population with SCD. Treatment options for sickle pain are limited and primarily involve the use of opioids. However, long-term opioid use is associated with numerous side effects. Thus, pain management in SCD remains a major challenge. Humanized transgenic mice expressing exclusively human sickle hemoglobin show features of pain and pathobiology similar to that in patients with SCD. Therefore, these mice offer the potential for investigating the mechanisms of pain in SCD and allow for development of novel targeted analgesic therapies. © 2018 by John Wiley & Sons, Inc.

Satellite Repeats Identify X Chromatin for Dosage Compensation in Drosophila melanogaster Males.

A common feature of sex chromosomes is coordinated regulation of X-linked genes in one sex. Drosophila melanogaster males have one X chromosome, whereas females have two. The resulting imbalance in gene dosage is corrected by increased expression from the single X chromosome of males, a process known as dosage compensation. In flies, compensation involves recruitment of the male-specific lethal (MSL) complex to X-linked genes and modification of chromatin to increase expression. The extraordinary selectivity of the MSL complex for the X chromosome has never been explained. We previously demonstrated that the small interfering RNA (siRNA) pathway and siRNA from a family of X-linked satellite repeats (1.688X repeats) promote X recognition. Now, we test the ability of 1.688X DNA to attract compensation to genes nearby and report that autosomal integration of 1.688X repeats increases MSL recruitment and gene expression in surrounding regions. Placement of 1.688X repeats opposite a lethal autosomal deletion achieves partial rescue of males, demonstrating functional compensation of autosomal chromatin. Females block formation of the MSL complex and are not rescued. The 1.688X repeats are therefore cis-acting elements that guide dosage compensation. Furthermore, 1.688X siRNA enhances rescue of males with a lethal deletion but only when repeat DNA is present on the intact homolog. We propose that the siRNA pathway promotes X recognition by enhancing the ability of 1.688X DNA to attract compensation in cis. The dense and near-exclusive distribution of 1.688X sequences along the X chromosome suggests that they play a primary role in determining X identity during dosage compensation.

Incidence of Postoperative Pain after Single Visit and Two Visit Root Canal Therapy: A Randomized Controlled Trial.

INTRODUCTION: Root Canal Treatment (RCT) has become a mainstream procedure in dentistry. A successful RCT is presented by absence of clinical signs and symptoms in teeth without any radiographic evidence of periodontal involvement. Completing this procedure in one visit or multiple visits has long been a topic of discussion. AIM: To evaluate the incidence of postoperative pain after root canal therapy performed in single visit and two visits. MATERIAL AND METHODS: An unblinded/ open label randomized controlled trial was carried out in the endodontic department of the Dental Institute, where 78 patients were recruited from the regular pool of patients. A total of 66 maxillary central incisors requiring root canal therapy fulfilled the inclusion and exclusion criteria. Using simple randomization by biased coin randomization method, the selected patients were assigned into two groups: group A (n=33) and group B (n=33). Single visit root canal treatment was performed for group A and two visit root canal treatment for group B. Independent sample t-test was used for statistical analysis. RESULTS: Thirty three patients were allotted to group A where endodontic treatment was completed in single visit while 33 patients were allotted to group B where endodontic treatment was completed in two visits. One patient dropped-out from Group A. Hence in Group A, 32 patients were analysed while in Group B, 33 patients were analysed. After 6 hours, 12 hours and 24 hours of obturation, pain was significantly higher in Group B as compared to Group A. However, there was no significant difference in the pain experienced by the patients 48 hours after treatment in both the groups. CONCLUSION: Incidence of pain after endodontic treatment being performed in one-visit or two-visits is not significantly different.

Modulation of Chromatin by Noncoding RNA.

Noncoding RNAs (ncRNAs) are remarkably powerful, flexible, and pervasive cellular regulators. The involvement of these molecules in virtually all aspects of eukaryotic chromatin function is notable. Long and short ncRNAs play broadly complementary roles in these processes. Short ncRNAs underlie a programmable system of chromatin modification that silences mobile elements, identifies boundaries, and initiates the formation of constitutive heterochromatin in yeast. In contrast, long noncoding RNAs (lncRNAs) enforce developmentally appropriate expression and switch gene expression programs. lncRNAs accomplish this through diverse mechanisms, but often by modulating the activity or localization of chromatin regulatory complexes. Both long and short ncRNAs play key roles in organization of complex genomes of higher eukaryotes, and their coordinated actions appear to underlie some of the more dramatic examples of epigenetic regulation. This review contrasts well-studied examples of chromatin regulation by RNA and introduces examples of coordination between these systems.

A strategy for generation and balancing of autosome: Y chromosome translocations.

We describe a method for generation and maintenance of translocations that move large autosomal segments onto the Y chromosome. Using this strategy we produced ( 2;Y) translocations that relocate between 1.5 and 4.8 Mb of the 2nd chromosome.. All translocations were easily balanced over a male-specific lethal 1 (msl-1) mutant chromosome. Both halves of the translocation carry visible markers, as well as P-element ends that enable molecular confirmation. Halves of these translocations can be separated to produce offspring with duplications and with lethal second chromosome deficiencies . Such large deficiencies are otherwise tedious to generate and maintain.

The effect of various surface contaminants on the microleakage of two different generation bonding agents: A stereomicroscopic study.

AIM: The aim of this in vitro study was to evaluate the microleakage of two different generation bonding agents in the presence of various surface contaminants. MATERIALS AND METHODS: Class V cavities were prepared on 150 extracted human permanent molars. The samples were randomly divided into two main groups of 75 teeth each. Group I: Fifth generation bonding system (Single Bond, 3M). Group II: Seventh generation bonding system (iBond, Kulzer). Subgroups were formed according to exposure to different surface contaminants (saliva, blood, caries disclosing agent and haemostatic agent). Cavities were restored with hybrid composite (Z-100, 3M) and evaluated for microleakage. The scores were subjected to 't' test and analysis of variance (ANOVA) test. RESULTS: Single Bond and iBond did not provide complete resistance to microleakage when there was no contamination. Microleakage was minimum in the no contamination subgroup and maximum with the haemostatic agent subgroup for both the groups. CONCLUSION: Single bond showed lesser micro leakage in contaminated conditions.

Reattachment of fractured anterior teeth-determining fracture strength using different techniques: an in vitro study.

AIM: The aim of this in vitro study was to evaluate and compare the shear bond strength of teeth reattached with sixth generation dentin bonding agent: Xeno III and microhybrid resin composite: Esthet-X, using three different techniques: (1) Simple reattachment, (2) overcontour and (3) internal dentinal groove. METHODOLOGY: A total of 70 human maxillary central incisors were selected and divided into four groups as follows. Group I: Control group comprised of 10 samples. Group II: Simple reattachment, group III: Overcontour and group IV: Internal dentinal groove. Groups II, III and IV comprised of 20 samples each. The teeth in three study groups were sectioned using a diamond disk and the fragment was reattached with Esthet-X and Xeno III using three different techniques. Specimens were stored in tap water for 24 hours and shear bond strength was determined using universal testing machine using a knife edge chisel (0.5 mm in cross-section) at a crosshead speed of 1 mm/minute. RESULTS: The results of this study showed following mean value of fracture strength in Kgf: Group I: Control-27.71; group II: Simple reattachment-9.78; group III: Overcontour-24.41; group IV: Internal dentinal groove-23.83. CONCLUSION: The overcontour technique had the highest strength recovery while the simple reattachment had the lowest. CLINICAL SIGNIFICANCE: The overcontour technique provided strength recovery almost similar to intact teeth emphasizing that tooth preparation influenced fracture resistance.

The effect of one-step and multi-step polishing systems on surface texture of two different resin composites.

OBJECTIVE: The purpose of this in-vitro study was to evaluate the surface roughness of two direct resin composites polished with one-step and multi-step polishing systems. MATERIALS AND METHODS: The resin composites examined in this study include minifill-hybrid composite Esthet-X (DENTSPLY/Caulk, Milford, DE, USA) and packable composite Solitaire II (Heraeus Kulzer, Inc., Southbend). A total of 42 discs (10 x 2 mm), 21 specimens of each restorative material were fabricated. Seven specimens per composite group received no polishing treatment and served as control. For each composite group, the specimens were randomly divided into two polishing systems: One-step PoGo (Dentsply/Caulk, Milford, DE, USA) and multi-step Super Snap (Shofu, Inc. Kyoto, Japan). Polishing systems were applied according to the manufacturer's instructions after being ground wet with 1200 grit silicon carbide paper. The surface roughness values were determined using a profilometer. RESULTS: Data was subjected to student's t test at a significance level of 0.05. The smoothest surfaces were achieved under Mylar strips in both the composite groups. Mean Ra values ranged from 0.09 to 0.3 mum for Esthet-X group and from 0.18 to 0.3 mum for Solitaire II with different polishing systems. The ranking of the order of surface roughness on the basis of the type of composite was as follows: Esthet-X < Solitaire II for PoGo system and Esthet-X = Solitaire II for Super Snap; and the ranking for the polishing system was: PoGo < Super Snap (P