Tamarind Seeds in Tracheobronchial Tree: Challenges during Rigid Bronchoscopy.

Background: Tamarind seed aspiration is not frequent in children and is usually observed in kids from rural backgrounds, with easy access to tamarind fruits and their by-products. Materials and Methods: We report a retrospective review of five patients managed in the pediatric surgery department for tamarind seed aspiration into the tracheobronchial tree. The data were analyzed based on age, clinical presentation, bronchoscopic observations, and the challenges faced during the rigid bronchoscopic retrieval and postoperative course. Results: There were four boys and one girl with a median age of 10 years. High-resolution computed tomography thorax was done in all patients. The foreign body was identified in the right main bronchus in one and the left main bronchus in four patients. All five patients underwent rigid bronchoscopy and retrieval of the seed. Two patients had an early presentation (within a week) - they needed temporary tracheotomy as the swollen seed could not be negotiated through the narrow glottis. Two patients had a late presentation (around 15 days) - they required removal in piecemeal using crushing forceps and multiple insertions of bronchoscope prolonging surgical time. One patient presented at 22 days posttamarind aspiration. It was soft enough for easy disintegration with crocodile forceps and expeditiously removed in three to four pieces. All patients recovered uneventfully. Conclusion: Removal of tamarind seed foreign body from the tracheobronchial tree is challenging. Anticipating the difficulties and being prepared well, helps to reduce the intraoperative difficulty, and allow successful removal with favorable patient outcomes.

Urothelial Carcinoma: Epidemiology and Imaging-Based Review.

Bladder cancer is the 6th most common malignancy in the United States, with urothelial carcinomas comprising over 95% of cases of bladder cancer, and commands a significant disease burden in Rhode Island. Imaging studies can provide valuable diagnostic information for urothelial carcinomas at initial presentation and are routinely used for noninvasive staging, treatment response monitoring, and post-treatment surveillance. This review aims to discuss and highlight three imaging modalities: ultrasonography, computed tomography, and magnetic resonance imaging, with particular focus on the notable features and appearance of urothelial carcinoma on each modality and their relative utility throughout the disease course. A general overview of disease epidemiology and treatment practices is also provided.

Proteomic cardiovascular risk assessment in chronic kidney disease.

AIMS: Chronic kidney disease (CKD) is widely prevalent and independently increases cardiovascular risk. Cardiovascular risk prediction tools derived in the general population perform poorly in CKD. Through large-scale proteomics discovery, this study aimed to create more accurate cardiovascular risk models. METHODS AND RESULTS: Elastic net regression was used to derive a proteomic risk model for incident cardiovascular risk in 2182 participants from the Chronic Renal Insufficiency Cohort. The model was then validated in 485 participants from the Atherosclerosis Risk in Communities cohort. All participants had CKD and no history of cardiovascular disease at study baseline when ∼5000 proteins were measured. The proteomic risk model, which consisted of 32 proteins, was superior to both the 2013 ACC/AHA Pooled Cohort Equation and a modified Pooled Cohort Equation that included estimated glomerular filtrate rate. The Chronic Renal Insufficiency Cohort internal validation set demonstrated annualized receiver operating characteristic area under the curve values from 1 to 10 years ranging between 0.84 and 0.89 for the protein and 0.70 and 0.73 for the clinical models. Similar findings were observed in the Atherosclerosis Risk in Communities validation cohort. For nearly half of the individual proteins independently associated with cardiovascular risk, Mendelian randomization suggested a causal link to cardiovascular events or risk factors. Pathway analyses revealed enrichment of proteins involved in immunologic function, vascular and neuronal development, and hepatic fibrosis. CONCLUSION: In two sizeable populations with CKD, a proteomic risk model for incident cardiovascular disease surpassed clinical risk models recommended in clinical practice, even after including estimated glomerular filtration rate. New biological insights may prioritize the development of therapeutic strategies for cardiovascular risk reduction in the CKD population.

Boosting In-Vivo Anti-Tumor Immunity with an Oral Microparticulate Breast Cancer Vaccine and Low-Dose Cyclophosphamide.

Tumor cells express antigens that should induce immune-mediated rejection; however, spontaneous rejection of established tumors is rare. Recent evidence suggests that patients suffering from cancer exhibit an elevation in regulatory T cells population, a subset of CD4+ T cells, which suppress tumor recognition and elimination by cytotoxic T cells. This study investigates immunotherapeutic strategies to overcome the immunosuppressive effects exerted by regulatory T cells. A novel immunotherapeutic strategy was developed by simultaneous administration of oral microparticulate breast cancer vaccines and cyclophosphamide, a regulatory T cell inhibitor. Breast cancer vaccine microparticles were prepared by spray drying, and administered orally to female mice inoculated with 4TO7 murine breast cancer cells in combination with a low dose of intraperitoneally administered cyclophosphamide. Mice receiving the combination of vaccine microparticles and cyclophosphamide exhibited maximal tumor regression and the highest survival rate compared with the control groups. This study highlights the importance of cancer vaccination along with regulatory T cell depletion in cancer therapy, and suggests that a low dose of cyclophosphamide that specifically and significantly depletes regulatory T cells may be a highly effective immunotherapeutic strategy for the treatment of cancer.

Enhanced Immunogenicity of Adjuvanted Microparticulate HPV16 Vaccines Administered via the Transdermal Route.

Human papillomavirus (HPV) causes cervical cancer among women and is associated with other anogenital cancers in men and women. Prophylactic particulate vaccines that are affordable, self-administered and efficacious could improve uptake of HPV vaccines world-wide. The goal of this research is to develop a microparticulate HPV16 vaccine for transdermal administration using AdminPatch® and assess its immunogenicity in a pre-clinical mouse model. HPV16 microparticles were prepared using a biocompatible polymer and characterized in terms of size, zeta potential, encapsulation efficiency and microparticle yield. Scanning and transmission electron microscopy were conducted to confirm particle image and to visualize the conformation of HPV16 vaccine particles released from microparticle formulation. In vivo studies performed to evaluate the potential of the microparticulate vaccine initiated a robust and sustained immune response. HPV16 IgG antibodies were significantly elevated in the microparticle group compared to antigen solutions administered by the transdermal route. Results show significant expansion of CD4+, CD45R, CD27 and CD62L cell populations in the vaccinated mice group, indicating the high efficacy of the microparticulate vaccine when administered via transdermal route. The findings of this study call attention to the use of minimally invasive, pain-free routes to deliver vaccine.

The Effect of Sealer Application Methods on Voids Volume after Aging of Three Calcium Silicate-Based Sealers: A Micro-Computed Tomography Study.

During obturation, air voids are undesirable as they may provide shelter for microorganisms or passage for fluids. This study aimed to compare the occurrence of voids between three calcium silicate-based sealers (CSBSs) (MTA-Fillapex, BioRoot-RCS, Bio-C) and the change in their volume after aging. In addition, we aimed to compare voids when using two sealer application methods: lentulo-spiral (LS) and gutta-percha (GP) cone. Thirty extracted mandibular premolars (n = 30) were endodontically prepared and obturated using single GP cone (SGPC) technique. Each sealer was applied to 10 teeth (n = 10) using LS or GP. Micro-computed tomography (micro-CT) was used to quantify the volume of root filling and voids before and after 8-week storage in a phosphate-rich medium. The percentage of root filling and voids were compared between the groups using a Mann-Whitney U test and Kruskal-Wallis test with a Bonferroni correction. Before aging, the percentages of root filling volume after obturation were comparable with no significant differences between sealers (p = 0.325) or application methods (p = 0.950). After aging, the voids' volume increased significantly in all sealers (p ≤ 0.05). However, no significant differences were found between sealers (p = 0.302). In conclusion, voids in CSBSs may not reduce in size with aging; hence, SGPC should be carefully selected for suitable cases.

The Bond Strength of a Universal Adhesive System with Silane to Lithium Disilicates in Comparison with an Isolated Silane Coupling Agent.

PURPOSE: The aim of this in vitro study was to evaluate the effects of different durations of silane coupling agent application compared to a universal adhesive system regarding the shear bond strength of two ceramic materials. MATERIALS AND METHODS: A total of 120 human molars were ground to the dentinal coronal third and then fixed into an acrylic resin holder. Lithium disilicate specimens were divided into two main groups according to the ceramic type: computer-aided design/computer-aided manufacturing IPS e.max CAD and heat-pressed Initial LiSi Press GC (dimensions of 4 × 3× 3 mm). Each main group was subdivided into 6 subgroups (n = 10) according to the duration of the silane and universal adhesive system application (20, 60, or 120 seconds) on the ceramic surface before cementation; then, the cementation procedures were performed. All specimens were subjected to 5000 thermal cycles at 5 and 55°C before testing. The shear bond strength was measured using a universal testing machine. ANOVA and Scheffe post hoc test multiple comparisons tests were conducted (α = 0.05). RESULTS: The shear bond strength increased as the duration of the silane and universal adhesive system application increased. The highest bond value for each material was found for the silane application at 120 seconds, with a significant difference between 120 and 60, and 20 seconds for both e. max CAD and Initial LiSi materials (p = 0.029 and p ˂ 0.001, respectively). No significant difference was found between 60 and 20 seconds when silane and universal adhesive system were applied for both e. max CAD and Initial LiSi materials (p = 0.169 and p = 0.120, respectively). All groups treated with the silane primer showed significantly higher values than the universal adhesive system for each application time (p ˂ 0.001). CONCLUSION: Increasing the duration of the silane coupling agent and universal adhesive system application to 120 seconds on the ceramic surface before cementation improved the shear bond strength of the ceramic-cement interface. Ceramic pretreatment with silane could be an essential step for bonding ceramic to dentin regardless of silane presence in the universal adhesive system.

Influence of Heat Treatment and Reinforcements on Tensile Characteristics of Aluminium AA 5083/Silicon Carbide/Fly Ash Composites.

The effect of reinforcements and thermal exposure on the tensile properties of aluminium AA 5083-silicon carbide (SiC)-fly ash composites were studied in the present work. The specimens were fabricated with varying wt.% of fly ash and silicon carbide and subjected to T6 thermal cycle conditions to enhance the properties through "precipitation hardening". The analyses of the microstructure and the elemental distribution were carried out using scanning electron microscopic (SEM) images and energy dispersive spectroscopy (EDS). The composite specimens thus subjected to thermal treatment exhibit uniform distribution of the reinforcements, and the energy dispersive spectrum exhibit the presence of Al, Si, Mg, O elements, along with the traces of few other elements. The effects of reinforcements and heat treatment on the tensile properties were investigated through a set of scientifically designed experimental trials. From the investigations, it is observed that the tensile and yield strength increases up to 160 °C, beyond which there is a slight reduction in the tensile and yield strength with an increase in temperature (i.e., 200 °C). Additionally, the % elongation of the composites decreases substantially with the inclusion of the reinforcements and thermal exposure, leading to an increase in stiffness and elastic modulus of the specimens. The improvement in the strength and elastic modulus of the composites is attributed to a number of factors, i.e., the diffusion mechanism, composition of the reinforcements, heat treatment temperatures, and grain refinement. Further, the optimisation studies and ANN modelling validated the experimental outcomes and provided the training models for the test data with the correlation coefficients for interpolating the results for different sets of parameters, thereby facilitating the fabrication of hybrid composite components for various automotive and aerospace applications.

Pattern of dental needs and advice on Twitter during the COVID-19 pandemic in Saudi Arabia.

AIM: To compare and evaluate the influence of the COVID-19 outbreak on tweets related to dental treatment needs and advice of Saudi Twitter users in 2020 by comparing them to the same time-period in 2019. METHODS: Eight independent searches based on dentistry related keywords: "teeth, mouth and gingiva" were carried out within the timeframe between the 23rd of March and the 21st of June for the years 2020 and 2019. Extracted tweets were analyzed by two calibrated examiners as tweets containing expressed dental needs and tweets for dental advice, while spam tweets were excluded. Descriptive analysis was performed to present the overview of the findings using SPSS. Bivariate analysis was performed with Pearson's Chi Square, Fisher's Exact test and Mann-Whitney U test. Statistical significance was set at p ≤ 0.05. RESULTS: A total of 595 tweets from the year 2019 and 714 tweets from the year 2020 were obtained. Overall, combined dental needs and advice tweets, retweets, likes, and replies were higher in 2020 compared to 2019. Dental needs tweets were higher in 2020 compared to 2019, while dental advice tweets were lower in 2020 compared to 2019. Statistically significant differences were found between 2020 and 2019 with regards to dental needs well as with dental advice (p < 0.05). In addition, statistically significant differences were found between 2019 and 2020 with presence of pain, urgency of the dental need and type of advisor (p < 0.05). CONCLUSION: An obvious impact of the pandemic can be seen in the form of increased self-reported dental needs, pain and urgency among the public in Saudi Arabia. This study highlights the importance of social media, specifically Twitter, in expressing the public needs and utilizing it as a platform for education and advice.

Dental Care in the Arab Countries During the COVID-19 Pandemic: An Infodemiological Study.

BACKGROUND: Twitter is a powerful platform which could be used to reflect on the demand and supply of dental services during a pandemic. The aim of this study was to examine the nature and dissemination of COVID-19 information related to dentistry on Twitter platform Arabic database during the COVID-19 pandemic. METHODOLOGY: One hundred and fifty independent searches with a combination of keywords for both COVID-19 and dentistry from a preselected Arabic keyword were carried out for the period from the 2nd of March (first confirmed cases of COVID-19) to the 6th of July 2020. Tweets were filtered to remove duplicate and unrelated tweets. The suitable tweets were 1,150. After calibration, two examiners coded the tweets following two main themes: COVID-19 and oral health-related information. Tweets were then compared with COVID-19 daily events in the Arab countries as reported by the World Health Organization (WHO). Descriptive analysis was performed to present the overview of the findings using Microsoft Excel. RESULTS: The most retweeted information was the help with urgent consultation or emergency dental treatment during COVID-19 tweeted by a dentist. There were 673 retweets and 1,116 likes of this tweet. The most common tweets related to oral health were needs of dental treatment (n=462, 39.5%) of which, toothaches or wisdom tooth problems constituted 48% of the related tweets. CONCLUSION: Based on the results of this study, it is obvious that social media users reacted to the COVID-19 threat to dental practices. Twitter as one of the social media platforms served as a connection between dental health professionals and patients.

Container Closure and Delivery Considerations for Intravitreal Drug Administration.

Intravitreal (IVT) administration of therapeutics is the standard of care for treatment of back-of-eye disorders. Although a common procedure performed by retinal specialists, IVT administration is associated with unique challenges related to drug product, device and the procedure, which may result in adverse events. Container closure configuration plays a crucial role in maintaining product stability, safety, and efficacy for the intended shelf-life. Careful design of primary container configuration is also important to accurately deliver small volumes (10-100 µL). Over- or under-dosing may lead to undesired adverse events or lack of efficacy resulting in unpredictable and variable clinical responses. IVT drug products have been traditionally presented in glass vials. However, pre-filled syringes offer a more convenient administration option by reducing the number of steps required for dose preparation there by potentially reducing the time demand on the healthcare providers. In addition to primary container selection, product development studies should focus on, among other things, primary container component characterization, material compatibility with the formulation, formulation stability, fill volume determination, extractables/leachables, and terminal sterilization. Ancillary components such as disposable syringes and needles must be carefully selected, and a detailed administration procedure that includes dosing instructions is required to ensure successful administration of the product. Despite significant efforts in improving the drug product and administration procedures, ocular safety concerns such as endophthalmitis, increased intraocular pressure, and presence of silicone floaters have been reported. A systematic review of available literature on container closure and devices for IVT administration can help guide successful product development.

Ongoing declines of woodland birds: Are restoration plantings making a difference?

Woodland birds are a species assemblage of conservation concern, and their persistence in fragmented agricultural landscapes is dependent on both the preservation of existing woodland remnants and the implementation of restoration plantings. However, little is known about the habitat-use and persistence of birds in fragmented agricultural landscapes. We present a detailed, population-oriented study of woodland birds in temperate eucalypt woodland restoration plantings and remnant woodland patches in the South-west Slopes bioregion of New South Wales, Australia. First, we undertook a 3-yr mark-recapture project to assess annual survival and site fidelity in restoration plantings and woodland remnants. We supplemented our recapture efforts with resightings of color-banded individuals. Second, we tracked individual birds of two species, Superb Fairywren (Malurus cyaneus) and Willie Wagtail (Rhipidura leucophrys), and documented snapshots of their home ranges and movement patterns during the breeding season. Annual survival in the woodland bird assemblage was lower than expected (51%). Home ranges of the Superb Fairywren were positively correlated with patch size, and were constrained by patch edges in linear sites. Superb Fairywrens and Willie Wagtails were more likely to travel longer distances between substrates while foraging in linear sites. Willie Wagtails engaged in significant gap-crossing (up to 400 m) between adjacent habitat patches. Our findings indicate that (1) patch isolation and certain patch configurations place resident birds at an energetic disadvantage, and (2) in our study area, woodland bird populations are continuing to decline. We recommend landscape-scale habitat restoration programs aim to address ongoing population declines. Studies such as ours conducted over longer time periods would provide a deeper understanding of habitat use and population processes of woodland birds in fragmented agricultural landscapes.

Impact of Kidney Function on the Blood Proteome and on Protein Cardiovascular Risk Biomarkers in Patients With Stable Coronary Heart Disease.

Background Chronic kidney disease (CKD) confers increased cardiovascular risk, not fully explained by traditional factors. Proteins regulate biological processes and inform the risk of diseases. Thus, in 938 patients with stable coronary heart disease from the Heart and Soul cohort, we quantified 1054 plasma proteins using modified aptamers (SOMAscan) to: (1) discern how reduced glomerular filtration influences the circulating proteome, (2) learn of the importance of kidney function to the prognostic information contained in recently identified protein cardiovascular risk biomarkers, and (3) identify novel and even unique cardiovascular risk biomarkers among individuals with CKD. Methods and Results Plasma protein levels were correlated to estimated glomerular filtration rate (eGFR) using Spearman-rank correlation coefficients. Cox proportional hazard models were used to estimate the association between individual protein levels and the risk of the cardiovascular outcome (first among myocardial infarction, stroke, heart failure hospitalization, or mortality). Seven hundred and nine (67.3%) plasma proteins correlated with eGFR at P<0.05 (ρ 0.06-0.74); 218 (20.7%) proteins correlated with eGFR moderately or strongly (ρ 0.2-0.74). Among the previously identified 196 protein cardiovascular biomarkers, just 87 remained prognostic after correction for eGFR. Among patients with CKD (eGFR <60 mL/min per 1.73 m2), we identified 21 protein cardiovascular risk biomarkers of which 8 are unique to CKD. Conclusions CKD broadly alters the composition of the circulating proteome. We describe protein biomarkers capable of predicting cardiovascular risk independently of glomerular filtration, and those that are prognostic of cardiovascular risk specifically in patients with CKD and even unique to patients with CKD.

Mechanisms of Sodium-Glucose Cotransporter 2 Inhibition: Insights From Large-Scale Proteomics.

OBJECTIVE: To assess the effects of empagliflozin, a selective sodium-glucose cotransporter 2 (SGLT2) inhibitor, on broad biological systems through proteomics. RESEARCH DESIGN AND METHODS: Aptamer-based proteomics was used to quantify 3,713 proteins in 144 paired plasma samples obtained from 72 participants across the spectrum of glucose tolerance before and after 4 weeks of empagliflozin 25 mg/day. The biology of the plasma proteins significantly changed by empagliflozin (at false discovery rate-corrected P < 0.05) was discerned through Ingenuity Pathway Analysis. RESULTS: Empagliflozin significantly affected levels of 43 proteins, 6 related to cardiomyocyte function (fatty acid-binding protein 3 and 4 [FABPA], neurotrophic receptor tyrosine kinase, renin, thrombospondin 4, and leptin receptor), 5 to iron handling (ferritin heavy chain 1, transferrin receptor protein 1, neogenin, growth differentiation factor 2 [GDF2], and ß2-microglobulin), and 1 to sphingosine/ceramide metabolism (neutral ceramidase), a known pathway of cardiovascular disease. Among the protein changes achieving the strongest statistical significance, insulin-like binding factor protein-1 (IGFBP-1), transgelin-2, FABPA, GDF15, and sulphydryl oxidase 2 precursor were increased, while ferritin, thrombospondin 3, and Rearranged during Transfection (RET) were decreased by empagliflozin administration. CONCLUSIONS: SGLT2 inhibition is associated, directly or indirectly, with multiple biological effects, including changes in markers of cardiomyocyte contraction/relaxation, iron handling, and other metabolic and renal targets. The most significant differences were detected in protein species (GDF15, ferritin, IGFBP-1, and FABP) potentially related to the clinical and metabolic changes that were actually measured in the same patients. These novel results may inform further studies using targeted proteomics and a prospective design.

Focal choroidal excavation and giant choroidal cavern in an eye with pachychoroid.

Focal choroidal excavation (FCE) is a localized excavation of the choroid, which can be diagnosed by enhanced depth optical coherence tomography (OCT). Choroidal caverns are focal cavitation areas in the choroid which appear hyporeflective on OCT. These are angular or round, empty spaces with posterior tail of hypertransmission. A 47-year-old female presented to us for a routine eye check-up. The best-corrected visual acuity was 20/20 in both the eyes. On fundus examination, a localized pigmented lesion was seen in the right eye inferior to the optic disc, while the fundus of the left eye was normal. Spectral-domain OCT scan through the lesion showed an FCE and a giant cavern with a posterior tail of hypertransmission. The diameter of the cavern was 977 µ ×264 µ, with a subfoveal choroidal thickness (SFCT) of 360 µ. The SFCT of the other eye was 300 µ. Coexistence of FCE and cavern in an eye with thickened choroid is not yet reported in the literature, and their coexistence provides possible insight into the formation of FCE and caverns in the setting of thickened choroid.

Harnessing T-cell activity against prostate cancer: A therapeutic microparticulate oral cancer vaccine.

Prostate Cancer specific immunotherapy in combination with immune stimulating adjuvants may serve as a viable strategy for facilitating tumor regression and preventing recurrence. In this study, an oral microparticulate vaccine encapsulating tumor associated antigens (TAA) extracted from a murine prostate cancer cell line, TRAMP-C2, was formulated with the help of a spray dryer. Microparticles were characterized in vitro to determine their physicochemical properties and antigenicity. Formulated microparticles had an average size of 4.92 ±â€¯0.5 µm with a zeta potential of 7.92 ±â€¯1.2 mV. In order to test our formulation for its ability to demonstrate adequate antigen presentation and co-stimulation, microparticles were tested in vitro on murine dendritic cells. In vitro biological characterization demonstrated the activation of specific immune system markers such as CD80/86, CD40, MHC-I and MHC-II. Following in vitro characterization, in vivo anti-tumor efficacy of the oral microparticulate vaccine was evaluated in C57BL/6 male mice. Combination therapy of vaccine microparticles with cyclophosphamide and granulocyte macrophage-colony stimulating factor (GM-CSF) demonstrated a five-fold reduction in tumor volume as compared to non-vaccinated mice. At the cellular level, cyclophosphamide and GM-CSF augmented the vaccine response as indicated by the reduced tumor volume and significant elevation of cytotoxic T-cell (CTL) CD8+ and (T-helper) CD4+ T-cells compared to mice receiving vaccine microparticles alone. Furthermore, our studies indicate a significant reduction in T-regulatory cells (T-regs) in mice receiving vaccine along with GM-CSF and cyclophosphamide, one of the immune escape mechanisms linked to tumor growth and progression. Thus, oral microparticulate vaccines have the potential to trigger a robust anti-tumor cellular response, and in combination with clinically relevant agents, significantly resist tumor growth and progression.

Improving Assessment of Drug Safety Through Proteomics: Early Detection and Mechanistic Characterization of the Unforeseen Harmful Effects of Torcetrapib.

BACKGROUND: Early detection of adverse effects of novel therapies and understanding of their mechanisms could improve the safety and efficiency of drug development. We have retrospectively applied large-scale proteomics to blood samples from ILLUMINATE (Investigation of Lipid Level Management to Understand its Impact in Atherosclerotic Events), a trial of torcetrapib (a cholesterol ester transfer protein inhibitor), that involved 15 067 participants at high cardiovascular risk. ILLUMINATE was terminated at a median of 550 days because of significant absolute increases of 1.2% in cardiovascular events and 0.4% in mortality with torcetrapib. The aims of our analysis were to determine whether a proteomic analysis might reveal biological mechanisms responsible for these harmful effects and whether harmful effects of torcetrapib could have been detected early in the ILLUMINATE trial with proteomics. METHODS: A nested case-control analysis of paired plasma samples at baseline and at 3 months was performed in 249 participants assigned to torcetrapib plus atorvastatin and 223 participants assigned to atorvastatin only. Within each treatment arm, cases with events were matched to controls 1:1. Main outcomes were a survey of 1129 proteins for discovery of biological pathways altered by torcetrapib and a 9-protein risk score validated to predict myocardial infarction, stroke, heart failure, or death. RESULTS: Plasma concentrations of 200 proteins changed significantly with torcetrapib. Their pathway analysis revealed unexpected and widespread changes in immune and inflammatory functions, as well as changes in endocrine systems, including in aldosterone function and glycemic control. At baseline, 9-protein risk scores were similar in the 2 treatment arms and higher in participants with subsequent events. At 3 months, the absolute 9-protein derived risk increased in the torcetrapib plus atorvastatin arm compared with the atorvastatin-only arm by 1.08% (P=0.0004). Thirty-seven proteins changed in the direction of increased risk of 49 proteins previously associated with cardiovascular and mortality risk. CONCLUSIONS: Heretofore unknown effects of torcetrapib were revealed in immune and inflammatory functions. A protein-based risk score predicted harm from torcetrapib within just 3 months. A protein-based risk assessment embedded within a large proteomic survey may prove to be useful in the evaluation of therapies to prevent harm to patients. CLINICAL TRIAL REGISTRATION: URL: https://www.clinicaltrials.gov. Unique identifier: NCT00134264.

Next-generation sequencing-based method shows increased mutation detection sensitivity in an Indian retinoblastoma cohort.

PURPOSE: Retinoblastoma (Rb) is the most common primary intraocular cancer of childhood and one of the major causes of blindness in children. India has the highest number of patients with Rb in the world. Mutations in the RB1 gene are the primary cause of Rb, and heterogeneous mutations are distributed throughout the entire length of the gene. Therefore, genetic testing requires screening of the entire gene, which by conventional sequencing is time consuming and expensive. METHODS: In this study, we screened the RB1 gene in the DNA isolated from blood or saliva samples of 50 unrelated patients with Rb using the TruSight Cancer panel. Next-generation sequencing (NGS) was done on the Illumina MiSeq platform. Genetic variations were identified using the Strand NGS software and interpreted using the StrandOmics platform. RESULTS: We were able to detect germline pathogenic mutations in 66% (33/50) of the cases, 12 of which were novel. We were able to detect all types of mutations, including missense, nonsense, splice site, indel, and structural variants. When we considered bilateral Rb cases only, the mutation detection rate increased to 100% (22/22). In unilateral Rb cases, the mutation detection rate was 30% (6/20). CONCLUSIONS: Our study suggests that NGS-based approaches increase the sensitivity of mutation detection in the RB1 gene, making it fast and cost-effective compared to the conventional tests performed in a reflex-testing mode.