Targeting DCAF5 suppresses SMARCB1-mutant cancer by stabilizing SWI/SNF.

Whereas oncogenes can potentially be inhibited with small molecules, the loss of tumour suppressors is more common and is problematic because the tumour-suppressor proteins are no longer present to be targeted. Notable examples include SMARCB1-mutant cancers, which are highly lethal malignancies driven by the inactivation of a subunit of SWI/SNF (also known as BAF) chromatin-remodelling complexes. Here, to generate mechanistic insights into the consequences of SMARCB1 mutation and to identify vulnerabilities, we contributed 14 SMARCB1-mutant cell lines to a near genome-wide CRISPR screen as part of the Cancer Dependency Map Project1-3. We report that the little-studied gene DDB1-CUL4-associated factor 5 (DCAF5) is required for the survival of SMARCB1-mutant cancers. We show that DCAF5 has a quality-control function for SWI/SNF complexes and promotes the degradation of incompletely assembled SWI/SNF complexes in the absence of SMARCB1. After depletion of DCAF5, SMARCB1-deficient SWI/SNF complexes reaccumulate, bind to target loci and restore SWI/SNF-mediated gene expression to levels that are sufficient to reverse the cancer state, including in vivo. Consequently, cancer results not from the loss of SMARCB1 function per se, but rather from DCAF5-mediated degradation of SWI/SNF complexes. These data indicate that therapeutic targeting of ubiquitin-mediated quality-control factors may effectively reverse the malignant state of some cancers driven by disruption of tumour suppressor complexes.

RI2AP: Robust and Interpretable 2D Anomaly Prediction in Assembly Pipelines.

Predicting anomalies in manufacturing assembly lines is crucial for reducing time and labor costs and improving processes. For instance, in rocket assembly, premature part failures can lead to significant financial losses and labor inefficiencies. With the abundance of sensor data in the Industry 4.0 era, machine learning (ML) offers potential for early anomaly detection. However, current ML methods for anomaly prediction have limitations, with F1 measure scores of only 50% and 66% for prediction and detection, respectively. This is due to challenges like the rarity of anomalous events, scarcity of high-fidelity simulation data (actual data are expensive), and the complex relationships between anomalies not easily captured using traditional ML approaches. Specifically, these challenges relate to two dimensions of anomaly prediction: predicting when anomalies will occur and understanding the dependencies between them. This paper introduces a new method called Robust and Interpretable 2D Anomaly Prediction (RI2AP) designed to address both dimensions effectively. RI2AP is demonstrated on a rocket assembly simulation, showing up to a 30-point improvement in F1 measure compared to current ML methods. This highlights its potential to enhance automated anomaly prediction in manufacturing. Additionally, RI2AP includes a novel interpretation mechanism inspired by a causal-influence framework, providing domain experts with valuable insights into sensor readings and their impact on predictions. Finally, the RI2AP model was deployed in a real manufacturing setting for assembling rocket parts. Results and insights from this deployment demonstrate the promise of RI2AP for anomaly prediction in manufacturing assembly pipelines.

Impact of self-assessment on dental student's performance in pre-clinical conservative dentistry course.

BACKGROUND: Self-assessment (SA) is an interactive course that endorses the accomplishment of learning objectives through learners' identification of insufficiencies in their didactic knowledge and pre-clinical skills. This study was planned to determine whether there is any improvement in the faculty assessment (FA) score following the implementation of SA in the Pre-clinical Conservative Dentistry Course. METHODS: Fifty-four first-semester dental students were given an introductory lecture followed by a demonstration for Class I Cavity Preparation in typhodont mandibular first molar. At the end of the demonstration, the Scoring Rubric (SR) was explained point-wise in the prepared cavities. During the next session, all students performed Class I cavity preparation and they were given an assessment sheet to enter their scores (SA1). All teeth were evaluated by the Grading Faculties in a blinded manner (FA1). Each participant was explained the difference in their respective SA1 from FA1 and their queries were resolved individually. During the next sessions, Students and Grading Faculties followed the same protocol and scores were recorded as SA2, FA2, SA3 and FA3. RESULTS: The mean score of SA1 was significantly higher than that of FA1 (p < 0.001). However, no significant difference was obtained between SA and FA in the second (p = 0.352) and third (p = 0.434) assessments. In contrast with first assessment, mean marks obtained in FA were higher compared to SA in both second and third assessments. There was a statistically significant improvement in mean marks obtained by the students over time (p < 0.001). CONCLUSION: SA endorsed student-faculty communication and enhanced student's poise and technical skills in operative pre-clinical dentistry.

Biomimetic approach to strengthen the incisal fracture composite build-up: an in vitro study.

OBJECTIVE: Incisal composite build-up shows a high failure susceptibility. The incorporation of fiber-reinforced composite (FRC) during composite restoration could improve its strength. Hence the study was planned to compare the effect of various positions of FRC on the strength of composite resin incisal build-ups. METHODS: In maxillary incisors (n = 90), 3 mm of the incisal edge was cut and teeth were categorized into three groups based on the location and number of fibers used during incisal composite build-up - Group I: composite resin; Group II: composite resin and a single fiber palatally and Group III: composite resin along with two fibers palatally. RESULTS: The data showed that group II had the maximum load-bearing values followed by group I and group III. CONCLUSION: Within the confines of our study, it can be concluded that the addition of FRC to the conventional incisal composite build-up increased the overall strength restoration. Such composite restoration reinforced with a single fiber on the palatal side showed the highest load-bearing capacity compared to two fibers reinforced and unreinforced composites. The common mode of failure in group I was in composite resin, in two fibers reinforced at fibers-composite junction, and in one fiber reinforced composite was in the remaining part of the tooth.

Comparative evaluation of effect of sodium hypochlorite and chlorhexidine in dental unit waterline on aerosolized bacteria generated during dental treatment.

BACKGROUND: In dentistry, nosocomial infection poses a great challenge to clinicians. The microbial contamination of water in dental unit waterlines (DUWLs) is ubiquitous. Such infected DUWLs can transmit oral microbes in the form of aerosols. Previous studies have suggested treating DUWLs with various disinfectants to reduce cross-contamination. The literature lacks a comparative evaluation of the effect of the use of 0.2% chlorhexidine (CHX) and 0.1% sodium hypochlorite (NaOCl) in DUWLs on aerosolized bacteria generated during dental procedures. OBJECTIVE: To compare the effect of NaOCl and CHX in DUWLs on aerosolized bacteria generated during restorative and endodontic procedures. MATERIALS AND METHODS: A total of 132 patients were equally divided into three groups (n = 44 in each group) according to the content of DUWL as follows. Group I-0.1% NaOCl Group II-0.2% CHX Group III-distilled water (Positive control) One-way ANOVA was performed and the Kruskal-Wallis test was used for intergroup comparison. RESULTS: For the restorative procedure, inter-group comparison of mean colony-forming units (CFU) scores showed a statistically significant difference between the groups (p - .001) with the score of group 3 higher than group 2 followed by group 1. For the endodontics, an inter-group comparison of CFU scores showed a statistically significant difference between the groups (p - .003) with the mean score in group 1 being the lowest and group 3 being the highest. CONCLUSION: The addition of NaOCl or CHX in DUWLs shows an effective reduction in aerosolized bacteria compared to distilled water.

Management of separated endodontic instruments: case reports.

Successful management of separated endodontic instruments is difficult, but several techniques are available for the retrieval of a separated instrument (SI). An appropriate treatment plan depends on various factors, including the anatomy and location of the tooth; the size, type, and location of the SI; and the skill of the clinician. However, the selection of the most appropriate management technique based on these factors can be a challenging decision that directly influences the success of the procedure. This article presents 4 symptomatic cases of SIs managed with various modalities as well as 1-year clinical and radiographic follow-up results. The management techniques applied in these cases included the use of ultrasonic tips, the wire loop method, safe-sided H-files, the braiding technique, and bypassing of the file. The medical history of the patients was noncontributory except for a single patient who had type 2 diabetes mellitus, which might have delayed healing. After instrument retrieval, the patients were evaluated clinically for relief of signs and symptoms as well as for their responses to various tests such as percussion, palpation, and tooth mobility. Radiographic evaluation at the 12-month follow-up was used to determine the periapical index. The clinical and radiographic results and patient-reported outcomes were satisfactory in all cases. The 4 cases presented demonstrate that the keys to successful management of an SI are understanding the factors that influence the case and selecting an appropriate therapeutic technique.

Med12 regulates ovarian steroidogenesis, uterine development and maternal effects in the mammalian egg.

The transcriptional factor MED12 is part of the essential mediator transcriptional complex that acts as a transcriptional coactivator in all eukaryotes. Missense gain-of-function mutations in human MED12 are associated with uterine leiomyomas, yet the role of MED12 deficiency in tumorigenesis and reproductive biology has not been fully explored. We generated a Med12 reproductive conditional knockout mouse model to evaluate its role in uterine mesenchyme, granulosa cells, and oocytes. Mice heterozygous for Med12 deficiency in granulosa cells and uterus (Med12fl/+ Amhr2-Cre) were subfertile, while mice homozygous for Med12 deficiency in granulosa cells and uterus (Med12fl/fl Amhr2-Cre) were infertile. Morphological and histological analysis of the Med12fl/fl Amhr2-Cre reproductive tract revealed atrophic uteri and hyperchromatic granulosa cells with disrupted expression of Lhcgr, Esr1, and Esr2. Med12fl/fl Amhr2-Cre mice estrous cycle was disrupted, and serum analysis showed blunted rise in estradiol in response to pregnant mare serum gonadotropin. Uterine atrophy was partially rescued by exogenous steroid supplementation with dysregulation of Notch1 and Smo expression in steroid supplemented Med12fl/fl Amhr2-Cre uteri, indicating intrinsic uterine defects. Oocyte-specific ablation of Med12 caused infertility without disrupting normal folliculogenesis and ovulation, consistent with maternal effects of Med12 in early embryo development. These results show the critical importance of Med12 in reproductive tract development and that Med12 loss of function does not cause tumorigenesis in reproductive tissues.

Osteopontin is linked with AKT, FoxO1, and myostatin in skeletal muscle cells.

INTRODUCTION: Osteopontin (OPN) polymorphisms are associated with muscle size and modify disease progression in Duchenne muscular dystrophy (DMD). We hypothesized that OPN may share a molecular network with myostatin (MSTN). METHODS: Studies were conducted in the golden retriever (GRMD) and mdx mouse models of DMD. Follow-up in-vitro studies were employed in myogenic cells and the mdx mouse treated with recombinant mouse (rm) or human (Hu) OPN protein. RESULTS: OPN was increased and MSTN was decreased and levels correlated inversely in GRMD hypertrophied muscle. RM-OPN treatment led to induced AKT1 and FoxO1 phosphorylation, microRNA-486 modulation, and decreased MSTN. An AKT1 inhibitor blocked these effects, whereas an RGD-mutant OPN protein and an RGDS blocking peptide showed similar effects to the AKT inhibitor. RMOPN induced myotube hypertrophy and minimal Feret diameter in mdx muscle. DISCUSSION: OPN may interact with AKT1/MSTN/FoxO1 to modify normal and dystrophic muscle. Muscle Nerve 56: 1119-1127, 2017.

Sparing of the dystrophin-deficient cranial sartorius muscle is associated with classical and novel hypertrophy pathways in GRMD dogs.

Both Duchenne and golden retriever muscular dystrophy (GRMD) are caused by dystrophin deficiency. The Duchenne muscular dystrophy sartorius muscle and orthologous GRMD cranial sartorius (CS) are relatively spared/hypertrophied. We completed hierarchical clustering studies to define molecular mechanisms contributing to this differential involvement and their role in the GRMD phenotype. GRMD dogs with larger CS muscles had more severe deficits, suggesting that selective hypertrophy could be detrimental. Serial biopsies from the hypertrophied CS and other atrophied muscles were studied in a subset of these dogs. Myostatin showed an age-dependent decrease and an inverse correlation with the degree of GRMD CS hypertrophy. Regulators of myostatin at the protein (AKT1) and miRNA (miR-539 and miR-208b targeting myostatin mRNA) levels were altered in GRMD CS, consistent with down-regulation of myostatin signaling, CS hypertrophy, and functional rescue of this muscle. mRNA and proteomic profiling was used to identify additional candidate genes associated with CS hypertrophy. The top-ranked network included α-dystroglycan and like-acetylglucosaminyltransferase. Proteomics demonstrated increases in myotrophin and spectrin that could promote hypertrophy and cytoskeletal stability, respectively. Our results suggest that multiple pathways, including decreased myostatin and up-regulated miRNAs, α-dystroglycan/like-acetylglucosaminyltransferase, spectrin, and myotrophin, contribute to hypertrophy and functional sparing of the CS. These data also underscore the muscle-specific responses to dystrophin deficiency and the potential deleterious effects of differential muscle involvement.

Multifaceted role of erlotinib in various cancer: nanotechnology intervention, patent landscape, and advancements in clinical trials.

Erlotinib (ELB) is a tyrosine kinase inhibitor that targets the activity of Epidermal Growth Factor Receptor (EGFR) protein found in both healthy and cancerous cells. It binds reversibly to the ATP-binding site of the EGFR tyrosine kinase. ELB was approved by the US Food and Drug Administration (FDA) in 2004 for advanced non-small cell lung cancer (NSCLC) treatment in patients who relapsed after at least one other therapy. It was authorized for use with gemcitabine in 2005 for the treatment of advanced pancreatic cancer. In addition to lung cancer, ELB has shown promising results in the treatment of other cancers, including breast, prostate, colon, pancreatic, cervical, ovarian, and head and neck cancers. However, its limited water solubility, as a BCS class II drug, presents biopharmaceutical problems. Nanoformulations have been developed to overcome these issues, including increased solubility, controlled release, enhanced stability, tumor accumulation, reduced toxicity, and overcoming drug resistance. In older patients, ELB management should involve individualized dosing based on age-related changes in drug metabolism and close monitoring for adverse effects. Regular assessments of renal and hepatic functions are essential. This review provides an overview of ELB's role of ELB in treating various cancers, its associated biopharmaceutical issues, and the latest developments in ELB-related nanotechnology interventions. It also covers ELB patents granted in previous years and the ongoing clinical trials.

Development of erlotinib-loaded nanotransferosomal gel for the topical treatment of ductal carcinoma in situ.

Aims: This study was aimed to formulate erlotinib (ERL)-loaded transferosomal gel (ERL@TG) intended for topical application for the treatment of ductal carcinoma in situ. Materials & methods: The optimized process involved a thin-film hydration method to generate ERL-loaded transferosomes (ERL@TFS), which was incorporated into a carbopol gel matrix to generate ERL@TG. The optimized formulation was characterized in vitro followed by cytotoxicity evaluation on MCF-7 breast cancer cell lines and acute toxicity and skin irritation studies was performed in vivo. Results: In a comparative assessment against plain ERL, ERL@TG displayed enhanced efficacy against MCF-7 cell lines, reflected in considerably lower IC50 values with an enhanced safety profile. Conclusion: Optimized ERL@TG was identified as a promising avenue for addressing ductal carcinoma in situ breast cancer.

Despite progress, breast cancer remains a significant cause of death. This study aimed to revolutionize the treatment of noninvasive ductal carcinoma in situ, a type of breast cancer, by developing a special gel that can be applied directly to the breast. The developed gel was in the nanoform, a 'nanotransfersomal' gel that contained erlotinib, a potent drug for breast cancer. To ensure its effectiveness, we evaluated the erlotinib-loaded transfersomal gel through various tests. The results confirmed that the gel was nano-sized and loaded with a high amount of erlotinib. Animal studies were conducted to check if the prepared gel caused any skin irritation and interestingly, there was no irritation observed on the animals' skin. Furthermore, we treated breast cancer cells with the developed gel using a method called MTT assay and the results showed improved cell-killing activity in comparison to plain drug. In conclusion, this special gel represents a breakthrough in breast cancer treatment. It offers hope for better outcomes in the fight against this disease. This innovative approach involves directly applying the gel on the affected area topically to increase patient compliance and decreasing side effects of drugs. This could potentially transform ductal carcinoma in situ breast cancer treatment, bringing us closer to improved treatments and outcomes.

Undergraduates' comparative assessment, utilizing a structured rubric, of hands-on versus video-supported presentation in class II cavity preparation.

BACKGROUND: Dental education is considered as a complex, challenging and often stressful educational procedure. Acquisition of psychomotor skills by undergraduate students is an important step in many health professions to become a successful professional. During under graduation, class II cavity preparation exercise is of utmost important in dentistry. OBJECTIVE: To compare class II cavities prepared by students after hands-on live demonstration and pre-recorded video demonstration using well-organised evaluation rubrics. METHOD: Preclinical dental students (n= 50) were divided into two groups. The students in group I (n= 25) attended a hands-on live demonstration performed by one faculty while students in group II (n= 25) watched a 15-minute pre-recorded procedural video on the projector. Both groups were appealed to prepare class II cavity for amalgam involving disto-occlusal surface of mandibular second molar articulated on jaw model (TRU LON study model, Jayna industries, Ghaziabad U.P., India). Following completion of the preparations, all teeth were collected, and labelled grades of prepared cavities were given according to prespecified rubrics. The data of scores were presented as means and standard deviation. Statistical analysis of data was executed using SPSS software. A paired t-test was used to compare scores between groups. RESULTS: The study shows that the video-supported demonstration of a cavity preparation was better than the live hands-on demonstration. A higher mean response for the procedural video group was found compared to the live demonstration group (p= 0.000133). CONCLUSION: Pre-recorded video-supported demonstration along with guidance by a tutor may be a viable alternative to hands-on live demonstration in cavity preparation procedures during undergraduate dental training. Moreover, rubric methods can be implemented in the teaching of various preclinical exercises for conservative dentistry and endodontics.

Comparative evaluation of four different root canal irrigation techniques for apical extrusion of sodium hypochlorite - An in vitro study.

Objective: This study compared the potential for apical extrusion of sodium hypochlorite (NaOCl) when using needle irrigation (NI), passive ultrasonic irrigation (PUI), passive subsonic irrigation (PSI), and negative pressure system (NP). Materials and Methods: One hundred freshly extracted human permanent anterior teeth with complete root formation were embedded rigidly in agarose gel-containing cresol purple. Teeth were randomly allocated to six groups: G1-NI; G2-PUI; G3-PSI; G4-NP; G5-positive control; and G6-negative control. The chemomechanical preparation was completed and 12 ml of 3% NaOCl was delivered for 180 s in all groups. Dye diffusion was standardized by doing gel photography after the NaOCl final irrigation the Chi-square test and Kruskal-Wallis one-way analysis of variance test were used for the data analysis. Results: G1 had statistically significant apical extrusion compared to G2 (P = 0.001), G3 (P = 0.001), and G4 (P = 0.001) groups. G4 showed the least amount of apical extrusion. Conclusions: pH-sensitive gel model is useful in evaluating NaOCl extrusion. NP produced least NaOCl extrusion as compared to the other three systems. Due consideration should be given to the potential for apical extrusion of the irrigant before the selection of an irrigation system.

Evaluation of aromatherapy on success rate of inferior alveolar nerve block in teeth with irreversible pulpitis: a prospective randomized clinical trial.

OBJECTIVE: This prospective, block randomized clinical trial was conducted to evaluate the effect of aromatherapy on the success rate of inferior alveolar nerve block (IANB) in teeth with irreversible pulpitis. METHOD AND MATERIALS: In this clinical trial, 46 patients fulfilling the inclusion criteria were randomly divided into two equal groups: group I (n = 22) received IANB in a closed operatory without any fragrance, whereas group II (n = 24) received IANB in a separate closed operatory saturated with lavender fragrance using a candle warmer. The modified dental anxiety scale (MDAS) for anxiety and visual analog scale (VAS) for pain were recorded preoperatively as well as during access cavity preparation. For MDAS, a total score of more than 18 was considered as tremendously anxious or dental phobic. No or mild pain on VAS was considered as success. Data were analyzed using paired t test and independent sample t test. P < .05 was considered as statistically significant. RESULTS: Difference in mean VAS (P = .749) and MDAS (P = 1.000) between both the groups was statistically nonsignificant. However there was a statistically significant difference in mean VAS (P = .000) and MDAS (P = .001) during access opening. CONCLUSION: Lavender aromatherapy can be used successfully to alleviate dental anxiety as well as to increase the anesthetic success rate of IANB in teeth with irreversible pulpitis.

The SWI/SNF complex in cancer - biology, biomarkers and therapy.

Cancer genome-sequencing studies have revealed a remarkably high prevalence of mutations in genes encoding subunits of the SWI/SNF chromatin-remodelling complexes, with nearly 25% of all cancers harbouring aberrations in one or more of these genes. A role for such aberrations in tumorigenesis is evidenced by cancer predisposition in both carriers of germline loss-of-function mutations and genetically engineered mouse models with inactivation of any of several SWI/SNF subunits. Whereas many of the most frequently mutated oncogenes and tumour-suppressor genes have been studied for several decades, the cancer-promoting role of mutations in SWI/SNF genes has been recognized only more recently, and thus comparatively less is known about these alterations. Consequently, increasing research interest is being focused on understanding the prognostic and, in particular, the potential therapeutic implications of mutations in genes encoding SWI/SNF subunits. Herein, we review the burgeoning data on the mechanisms by which mutations affecting SWI/SNF complexes promote cancer and describe promising emerging opportunities for targeted therapy, including immunotherapy with immune-checkpoint inhibitors, presented by these mutations. We also highlight ongoing clinical trials open specifically to patients with cancers harbouring mutations in certain SWI/SNF genes.

Bilateral advanced (group D or E) intraocular retinoblastoma: outcomes in 72 Asian Indian patients.

PURPOSE: To study the clinical presentation, treatment, and outcomes of patients with bilateral advanced intraocular retinoblastoma. METHODS: Retrospective case series of 72 patients. RESULTS: The mean age at presentation was 19 months. Leukocoria (n = 49, 68%) was the most common presenting complaint. The tumors were classified as groups D (n = 60, 42%) or E (n = 84, 58%) based on the Philadelphia version of International Classification of Retinoblastoma (ICRB); groups D (n = 84, 58%) or E (n = 60, 42%) based on Children's Hospital Los Angeles version of International Classification of Intraocular Retinoblastoma (ICIoR); T2 (n = 116, 81%) or T3 (n = 28, 19%) based on 8th edition American Joint Committee Classification (AJCC). Systemic chemotherapy (n = 138, 96%) was the most common primary treatment modality. The chance of globe salvage was higher for group D based on ICRB (83%; odds ratio (OR) 7.73; 95% confidence interval (CI) 3.45-17.33) or ICIoR (81%; OR 12.75; 95% CI 5.74-28.34) and T2b (73%; OR 5.19; 95% CI 2.51-10.73) based on AJCC. Over a mean follow-up period of 59 months, tumor recurrence was noted in 42 (29%) eyes and globe salvage was achieved in 83 (58%) eyes. Of the 50 eyes where vision was recorded, vision of 20/200 or better was achieved in 24 (48%) eyes. There were events of leukemia (n = 1, 1%), pinealoblastoma (n = 1, 1%), systemic metastasis (n = 3, 4%), and death (n = 4, 6%) during the follow-up period. CONCLUSION: Multimodality treatment allows globe salvage (58%) and vision salvage (48%) in eyes with advanced group D and E intraocular retinoblastoma.

Accuracy of Different Apex Locators in Teeth with Simulated Apical Root Resorption: an In Vitro Study.

BACKGROUND: Accurate working length (WL) determination is necessary in achieving optimal healing by non-surgical root canal therapy in teeth with inflammatory apical root resorption. Electronic apex locators (EALs) are one of the mainstays in determination of WL of teeth. AIM: This study evaluated the accuracy of three EALs [RootZX (third generation), iPex (fourth generation) and Raypex 6 (modification of a fifth generation)] in determining the WL of teeth with simulated apical root resorption in permanent teeth. MATERIALS AND METHODS: Forty freshly extracted maxillary anterior teeth were collected and a 45° oblique cut was made at the root apex with a disc to simulate apical root resorption. Actual working length (AWL) was determined by direct visual method and was used as a control. Electronic working length (EWL) values were measured by three different apex locators that are RootZX (Group I), iPex (Group II) and Raypex 6 (Group III) at apex, 0.5 mm and 1 mm from apex. All values obtained were tabulated and statistical evaluation was carried out. RESULTS: At apex, EWL obtained using iPex (p=0.05) showed a statistically significant difference from AWL. At 0.5 mm and 1.0 mm tolerance, iPex showed non-acceptability for WL measurement in 67.5% and 17.5% of samples compared to Root ZX (12.5% and 2.5%) and Raypex (7.5% and none) respectively. CONCLUSION: Within the limitation of this study, it can be concluded that Raypex 6 and RootZX show statistically significant accuracy in WL measurement compared with iPex in teeth with apical root resorption.

Highly heterogeneous genomic landscape of uterine leiomyomas by whole exome sequencing and genome-wide arrays.

OBJECTIVE: To determine the genomic signatures of human uterine leiomyomas and prevalence of MED12 mutations in human uterine leiomyosarcomas. DESIGN: Retrospective cohort study. SETTING: Not applicable. PATIENT(S): This study included a set of 16 fresh frozen leiomyoma and corresponding unaffected myometrium specimens as well as 153 leiomyosarcomas collected from women diagnosed with uterine leiomyomas or leiomyosarcomas who underwent clinically indicated abdominal hysterectomy. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Whole exome sequencing and high-resolution X-chromosome and whole genome single nucleotide polymorphism microarray analyses were performed on leiomyoma samples negative for the known MED12 mutations and compared with their corresponding myometrium. Leiomyosarcoma specimens were examined for exon 2 MED12 mutations to evaluate the frequency of MED12 mutated leiomyosarcomas. RESULT(S): Our results indicate remarkable genomic heterogeneity of leiomyoma lesions. MED12-negative leiomyomas contain copy number alterations involving the Mediator complex subunits such as MED8, MED18, CDK8, and long intergenic nonprotein coding RNA340 (CASC15), which may affect the Mediator architecture and/or its transcriptional activity. We also identified mutations in a number of genes that were implicated in leiomyomagenesis such as COL4A6, DCN, and AHR, as well as novel genes: NRG1, ADAM18, HUWE1, FBXW4, FBXL13, and CAPRIN1. CONCLUSION(S): Mutations in genes implicated in cell-to-cell interactions and remodeling of the extracellular matrix and genomic aberrations involving genes coding for the Mediator complex subunits were identified in uterine leiomyomas. Additionally, we discovered that ∼4.6% of leiomyosarcomas harbored MED12 exon 2 mutations, but the relevance of this association with molecular pathogenesis of leiomyosarcoma remains unknown.

Anaesthesia Techniques for Maxillary Molars - A Questionnaire-Based Retrospective Field Survey of Dentist in Western India.

INTRODUCTION: Clinicians use various anaesthesia techniques like Posterior Superior Alveolar (PSA) nerve block, buccal infiltration with or without supplemental anaesthesia like palatal and intraligamentary infiltrations for root canal treatment in maxillary molars. However there is no general consensus regarding which technique is enough for performing endodontic treatment in maxillary molars. AIM: The aim of this questionnaire-based survey is to compare and evaluate the various techniques used to anaesthetize the maxillary molars and its effect on postoperative pain. MATERIALS AND METHODS: The data were obtained from 290 dental practitioners using a specially prepared questionnaire survey conducted anonymously. The questionnaire contained questions covering data such as years in dentistry, acquired specialty, techniques used for anaesthetizing maxillary molars, success of anaesthesia, and postoperative pain, etc. RESULTS: Buccal infilteration with supplemental anaesthesia in the form of palatal (82%) and intra-ligamentary infilteration (88%) show higher success rate compared to only buccal infilteration (69%). However, intra-ligamentary infilteration group showed highest rate (75%) of postoperative pain. General practitioners (62% of clinicians) prefer to give both buccal and palatal infilterations and specialists opt for only buccal infilteration (66-74% of specialists). CONCLUSION: Only buccal infilteration is sufficient during root canal treatment of maxillary molars. Routine use of supplemental anaesthesia in the form of palatal and intra-ligamentary infilteration is not necessary unless patient experiences discomfort during endodontic treatment. However, intra-ligamentary infilteration may lead to postoperative discomfort in the form of pain.