Ehmt2 inactivation in pancreatic epithelial cells shapes the transcriptional landscape and inflammation response of the whole pancreas.

Introduction: The Euchromatic Histone Methyl Transferase Protein 2 (EHMT2), also known as G9a, deposits transcriptionally repressive chromatin marks that play pivotal roles in the maturation and homeostasis of multiple organs. Recently, we have shown that Ehmt2 inactivation in the mouse pancreas alters growth and immune gene expression networks, antagonizing Kras-mediated pancreatic cancer initiation and promotion. Here, we elucidate the essential role of Ehmt2 in maintaining a transcriptional landscape that protects organs from inflammation. Methods: Comparative RNA-seq studies between normal postnatal and young adult pancreatic tissue from Ehmt2 conditional knockout animals (Ehmt2 fl/fl ) targeted to the exocrine pancreatic epithelial cells (Pdx1-Cre and P48 Cre/+ ), reveal alterations in gene expression networks in the whole organ related to injury-inflammation-repair, suggesting an increased predisposition to damage. Thus, we induced an inflammation repair response in the Ehmt2 fl/fl pancreas and used a data science-based approach to integrate RNA-seq-derived pathways and networks, deconvolution digital cytology, and spatial transcriptomics. We also analyzed the tissue response to damage at the morphological, biochemical, and molecular pathology levels. Results and discussion: The Ehmt2 fl/fl pancreas displays an enhanced injury-inflammation-repair response, offering insights into fundamental molecular and cellular mechanisms involved in this process. More importantly, these data show that conditional Ehmt2 inactivation in exocrine cells reprograms the local environment to recruit mesenchymal and immunological cells needed to mount an increased inflammatory response. Mechanistically, this response is an enhanced injury-inflammation-repair reaction with a small contribution of specific Ehmt2-regulated transcripts. Thus, this new knowledge extends the mechanisms underlying the role of the Ehmt2-mediated pathway in suppressing pancreatic cancer initiation and modulating inflammatory pancreatic diseases.

EHMT2 Inactivation in Pancreatic Epithelial Cells Shapes the Transcriptional Landscape and Inflammation Response of the Whole Pancreas.

The Euchromatic Histone Methyl Transferase Protein 2 (EHMT2), also known as G9a, deposits transcriptionally repressive chromatin marks that play pivotal roles in the maturation and homeostasis of multiple organs. Recently, we have shown that EHMT2 inactivation alters growth and immune gene expression networks, antagonizing KRAS-mediated pancreatic cancer initiation and promotion. Here, we elucidate the essential role of EHMT2 in maintaining a transcriptional landscape that protects organs from inflammation. Comparative RNA-seq studies between normal postnatal and young adult pancreatic tissue from EHMT2 conditional knockout animals ( EHMT2 fl/fl ) targeted to the exocrine pancreatic epithelial cells ( Pdx1-Cre and P48 Cre/+ ), reveal alterations in gene expression networks in the whole organ related to injury-inflammation-repair, suggesting an increased predisposition to damage. Thus, we induced an inflammation repair response in the EHMT2 fl/fl pancreas and used a data science-based approach to integrate RNA-seq-derived pathways and networks, deconvolution digital cytology, and spatial transcriptomics. We also analyzed the tissue response to damage at the morphological, biochemical, and molecular pathology levels. The EHMT2 fl/fl pancreas displays an enhanced injury-inflammation-repair response, offering insights into fundamental molecular and cellular mechanisms involved in this process. More importantly, these data show that conditional EHMT2 inactivation in exocrine cells reprograms the local environment to recruit mesenchymal and immunological cells needed to mount an increased inflammatory response. Mechanistically, this response is an enhanced injury-inflammation-repair reaction with a small contribution of specific EHMT2-regulated transcripts. Thus, this new knowledge extends the mechanisms underlying the role of the EHMT2-mediated pathway in suppressing pancreatic cancer initiation and modulating inflammatory pancreatic diseases.

Analyzing Instagram Posts on Hypothyroidism: Characteristics, Information Types, Quality, and Reliability.

Background and aims In the age of social media, a vast amount of information is widely and easily accessible. Platforms such as Instagram allow its users to post pictures and videos that can reach millions of users. This could be utilized by healthcare providers to provide education to a vast number of the population about a disease such as hypothyroidism with an easily digestible infographic. However, this easy accessibility comes with the risk of rampant misinformation. This study aimed to evaluate the characteristics of Instagram posts, the type of information, and the quality and reliability of information posted about hypothyroidism. Methodology This is a cross-sectional observational study that was conducted over the course of days on Instagram. Top posts meeting inclusion criteria under seven different hypothyroidism-related hashtags were surveyed for content and social media metrics by the authors utilizing Google Forms. The quality and reliability of the posts were analyzed using the global quality scale and DISCERN scales, respectively. The data were exported to an Excel sheet and analyzed using the SPSS software version 21.0 (Armonk, NY: IBM Corp.). Results A total of 629 posts met the inclusion criteria of which 62.5% were images and 37.5% were reels. The content heavily focused on the medical aspect of hypothyroidism with posts about symptoms (46.1%), prevention (39.59%), cause/etiology (36.41%), and treatment (34.34%). The median DISCERN score which reflects the reliability of the posts uploaded was highest for doctors at 3 and the least reliable posts were uploaded by dieticians, naturopathic doctors, and patients. This study found that the quality of posts uploaded by nutritionists and naturopathic doctors with a median Global Quality Score (GQS) score of 3. Conclusions There is a need to establish a quality control body that regulates the quality and reliability of the posts to curb misinformation and help patients gain easy access to reliable resources that will aid their decision-making.

Unraveling Abdominal Migraine in Adults: A Comprehensive Narrative Review.

Abdominal migraine is a condition characterized by recurrent episodes of abdominal pain accompanied by migraine-associated symptoms, primarily affecting pediatric populations. Its occurrence in adults is often overlooked due to limited literature on adult abdominal migraine. This article provides an overview of the current understanding and management of abdominal migraine in adult populations, including the diagnostic criteria, pathophysiology, differentiating features of other associated gastrointestinal pain syndromes, and various treatment approaches based on available literature. The review acknowledges the limitations, including the scarcity of literature on adult abdominal migraine and the absence of a systematic approach. It emphasizes the need for further research to enhance our understanding of this condition and establish evidence-based treatment guidelines specifically for adults. Accurate diagnosis and patient education are crucial for physicians in recognizing abdominal migraine as a differential diagnosis in cases of long-standing recurrent abdominal pain, promoting the importance of further research to advance our knowledge and improve patient outcomes.

Incompatibility of Pure SnO2 Thin Films for Room-Temperature Gas Sensing Application.

Despite the high sensitivity and selectivity, the high operating temperature required for activation energy of tin oxide (SnO2) still stands as a drawback for SnO2 based gas sensors. In this work, the SnO2 thin films were deposited through spray pyrolysis and were subjected to gas sensing at 27 °C (room temperature) towards different gases. The films exhibited a consistently low response of approximately 1 when tested to various VOCs. The type, concentration, and mobility of charge carriers were determined from the Hall measurements. The high carrier concentration accompanied by poor mobility and grain boundary scattering is supposed to hinder its performance at room temperature. The obtained film had spherical morphology, which lead to grain boundary scatterings and decreased the mobility of carriers.

Geometric morphometric analysis for sex determination using lateral cephalograms in Indian population: A preliminary study.

BACKGROUND: Orthodontic science centers primarily on the growth, development and advancement of the craniofacial structures. Geometric morphometrics (GMM) is a new approach for shape identification in forensic sciences. PURPOSE: The aim of this study was to examine the shape variation in the Indian sample in order to assess sexual dimorphism by application of two-dimensional GMM. MATERIALS AND METHODS: The sample comprised a total of 105 lateral cephalograms (54 males-51 females) of Angle's Class I malocclusion patients that were later subjected to principal component (PC) analysis and discriminant analysis. RESULTS: The PC analysis showed over 96% of shape variation. The initial three PCs were statistically significant that depicted as 58.37% of total shape variability, with PC1 represent for the most significant variance 28.48%, PC2 described 18.83% and PC3 11.06%. CONCLUSION: Sex of an individual was clearly associated with occlusion of teeth and showed considerable variation. GMM is an alternative research tool and can be utilized for diagnosing individual characterization and classification of malocclusion.

A novel multiscale and multipath convolutional neural network based age-related macular degeneration detection using OCT images.

BACKGROUND AND OBJECTIVE: One of the significant retinal diseases that affected older people is called Age-related Macular Degeneration (AMD). The first stage creates a blur effect on vision and later leads to central vision loss. Most people overlooked the primary stage blurring and converted it into an advanced stage. There is no proper treatment to cure the disease. So the early detection of AMD is essential to prevent its extension into the advanced stage. This paper proposes a novel deep Convolutional Neural Network (CNN) architecture to automate AMD diagnosis early from Optical Coherence Tomographic (OCT) images. METHODS: The proposed architecture is a multiscale and multipath CNN with six convolutional layers. The multiscale convolution layer permits the network to produce many local structures with various filter dimensions. The multipath feature extraction permits CNN to merge more features regarding the sparse local and fine global structures. The performance of the proposed architecture is evaluated through ten-fold cross-validation methods using different classifiers like support vector machine, multi-layer perceptron, and random forest. RESULTS: The proposed CNN with the random forest classifier gives the best classification accuracy results. The proposed method is tested on data set 1, data set 2, data set 3, data set 4, and achieved an accuracy of 0.9666, 0.9897, 0.9974, and 0.9978 respectively, with random forest classifier. Also, we tested the combination of first three data sets and achieved an accuracy of 0.9902. CONCLUSIONS: An efficient algorithm for detecting AMD from OCT images is proposed based on a multiscale and multipath CNN architecture. Comparison with other approaches produced results that exhibit the efficiency of the proposed algorithm in the detection of AMD. The proposed architecture can be applied in rapid screening of the eye for the early detection of AMD. Due to less complexity and fewer learnable parameters.

In situ thermosensitive hybrid mesoporous silica: preparation and the catalytic activities for carbonyl compound reduction.

In this study, free-radical polymerisation inside MCM-41 mesopores was examined to expose a construction route for a temperature-responsive switchable polymer-silica nanohybrid material with well-defined porosity. Herein, we introduced a vinyl monomer (N-isopropyl acrylamide), a cross-linker, and an AIBN initiator into the palladium nanoparticle incorporated MCM-41 pore channels using the wet-impregnation method followed by in situ radical polymerisation. The structural properties of the synthesised PNIPAM-PdNP-MCM-41 catalyst were analysed by various sophisticated analytical techniques. The temperature switchable nanohybrid catalyst was used to reduce carbonyl compounds to their corresponding alcohols. The catalyst showed high catalytic efficiency and robustness in an aqueous medium at 25 °C. Moreover, the system's polymer layer remarkably boosted catalytic selectivity and activity for carbonyl compound reduction as compared to other controlled catalysts. The suggested switchable system can be employed as a temperature-controllable heterogeneous catalyst and highlights a substitute technique to counter the methodical insufficiency in switchable supported molecular catalytic system production.

Synthesis of size-controlled and highly monodispersed silica nanoparticles using a short alkyl-chain fluorinated surfactant.

Highly monodispersed silica nanoparticles (SiNPs) were synthesised using a fluorinated surfactant, HOCH2CH(CF3)CO2H, and its efficiency was compared with efficiencies of five other surfactants. The size of the SiNPs (∼50-200 nm) was controlled by controlling the surfactant amount. The short alkyl-chain fluoro surfactant was found to be more efficient at producing monodispersed SiNPs than its long alkyl-chain fluoro or non-fluorinated surfactant counterparts.

RPE layer detection and baseline estimation using statistical methods and randomization for classification of AMD from retinal OCT.

BACKGROUND AND OBJECTIVE: Age-related macular degeneration (AMD) is a condition of the eye that affects the aged people. Optical coherence tomography (OCT) is a diagnostic tool capable of analyzing and identifying the disease affected retinal layers with high resolution. The objective of this work is to extract the retinal pigment epithelium (RPE) layer and the baseline (natural eye curvature, particular to every patient) from retinal spectral-domain OCT (SD-OCT) images. It uses them to find the height of drusen (abnormalities) in the RPE layer and classify it as AMD or normal. METHODS: In the proposed work, the contrast enhancement based adaptive denoising technique is used for speckle elimination. Pixel grouping and iterative elimination based on the knowledge of typical layer intensities and positions are used to obtain the RPE layer. Using this estimate, randomization techniques are employed, followed by polynomial fitting and drusen removal to arrive at a baseline estimate. The classification is based on the drusen height obtained by taking the difference between the RPE and baseline levels. We have used a patient, wise classification approach where a patient is classified diseased if more than a threshold number of patient images have drusen of more than a certain height. Since all slices of an affected patient will not show drusen, we are justified in adopting this technique. RESULTS: The proposed method is tested on a public data set of 2130 images/slices, which belonged to 30 patient volumes (15 AMD and 15 Normal) and achieved an overall accuracy of 96.66%, with no false positives. In comparison with existing works, the proposed method achieved higher overall accuracy and a better baseline estimate. CONCLUSIONS: The proposed work focuses on AMD/normal classification using a statistical approach. It does not require any training. The proposed method modifies the motion restoration paradigm to obtain an application-specific denoising algorithm. The existing RPE detection algorithm is modified significantly to make it robust and applicable even to images where the RPE is not very evident/there is a significant amount of perforations (drusen). The baseline estimation algorithm employs a powerful combination of randomization, iterative polynomial fitting, and pixel elimination in contrast to mere fitting techniques. The main highlight of this work is, it achieved an exact estimation of the baseline in the retinal image compared to the existing methods.

Dual Stimuli-Responsive Copper Nanoparticles Decorated SBA-15: A Highly Efficient Catalyst for the Oxidation of Alcohols in Water.

In the present work, a temperature and pH-responsive hybrid catalytic system using copolymer-capped mesoporous silica particles with metal nanoparticles is proposed. The poly(2-(dimethylamino)ethyl methacrylate)(DMAEMA)-co-N-tert-butyl acrylamide) (TBA)) shell on mesoporous silica SBA-15 was obtained through free radical polymerization. Then, copper nanoparticles (CuNPs) decorated SBA-15/copolymer hybrid materials were synthesized using the NaBH4 reduction method. SBA-15 was functionalized with trimethoxylsilylpropyl methacrylate (TMSPM) and named TSBA. It was found that the CuNPs were uniformly dispersed in the mesoporous channels of SBA-15, and the hybrid catalyst exhibited excellent catalytic performance for the selective oxidation of different substituted benzyl alcohols in water using H2O2 as an oxidant at room temperature. The dual (temperature and pH-) responsive behaviors of the CuNPs/p(DMAEMA-co-TBA)/TSBA catalyst were investigated using the dynamic light scattering technique. The conversion of catalytic products and selectivity were calculated using gas chromatographic techniques, whereas the molecular structure of the products was identified using 1H and 13C nuclear magnetic resonance (NMR) spectroscopy. The catalyst showed excellent catalytic activity toward the oxidation of alcohol to aldehyde in an aqueous medium below the lower critical solution temperature (LCST) and pKa values (7-7.5) of the copolymer. The main advantages of the hybrid catalyst, as compared to the existing catalysts, are outstanding alcohol conversion (up to 99%) for a short reaction time (1 h), small amount of the catalyst (5 mg), and good recyclability equal to at least five times.

Measurement of non-invasive blood pressure in lateral decubitus position under general anaesthesia - Which arm gives more accurate BP in relation to invasive BP - dependent or non-dependent arm?

BACKGROUND AND AIMS: Non-invasive blood pressure (NiBP) varies with the arm and body position. In the lateral decubitus position (LDP), the non-dependent arm reads lower, and the dependent arm reads higher pressure. We aimed to study the correlation between the NiBP and invasive arterial blood pressure (ABP) as anaesthesia progressed and its correlation in different BP ranges. METHODS: American Society of Anesthesiologists (ASA I-III) patients, between 18-70 years undergoing neurosurgical procedures in the LDP were studied. All were anaesthetised using a standard protocol, positioned in the LDP. NiBP was measured every 15 min in both dependent and non-dependent arms and correlated with the ABP. RESULTS: Intra-class correlation (ICC) done between the dependent arm NiBP and ABP showed good correlation for mean and systolic BP and moderate correlation for diastolic BP. ICC was 0.800, 0.846 and 0.818 for mean and 0.771, 0.782, 0.792 for systolic BP at 15 min, 1 h, and 2 h, respectively. The ICC between the non-dependent arm NiBP and the invasive ABP showed poor correlation for all BP (systolic, diastolic and mean). As anaesthesia progressed, the mean difference between the NiBP and the ABP decreased in the dependent arm and increased in the non-dependent arm. The strength of agreement between the NiBP and the ABP in various BP ranges showed moderate correlation for the dependent arm NiBP (0.45-0.54) and poor correlation (0.21-0.38) for the non-dependent arm. CONCLUSION: The NiBP of the dependent arm correlated well with ABP in LDP under general anaesthesia (GA). It is better to defer measuring NiBP in the non-dependent arm as the correlation with ABP is poor.

Palladium nanoparticles-anchored dual-responsive SBA-15-PNIPAM/PMAA nanoreactor: a novel heterogeneous catalyst for a green Suzuki-Miyaura cross-coupling reaction.

To develop a sustainable and cost-effective catalyst for cross-coupling reactions, dual (temperature and pH)-responsive poly(N-isopropyl acrylamide-co-methacrylic acid) (PNIPAM/PMAA) functionalised SBA-15 was synthesised via free radical polymerisation using potassium persulfate as an initiator and decorated with palladium nanoparticles (PdNPs-SBA-15-PNIPAM/PMAA). The X-ray photoelectron spectroscopic analysis revealed that the Pd content in the zero oxidation state of the catalyst was 1.21 wt%. The dynamic light scattering studies showed that the catalyst exhibited swelling behaviours at low temperatures (<32 °C) and high pH (>4), but exhibited deswelling behaviours at high temperatures (>32 °C) and low pH (<4). To examine the performance of the catalyst, Suzuki-Miyaura cross-coupling (SMC) reaction was conducted under batch reaction conditions. The reaction conditions were optimised with various parameters using phenylboronic acid and bromobenzene as the model substrates. High conversions (>90%) were realized for the room-temperature SMC reaction in an aqueous medium for various substituted aryl halides, while the conversion was low at relatively high temperatures (>32 °C). The conversion was dependent on the different electronic effects between the electron-releasing and electron-withdrawing groups of the aryl halides. After the experiment, the catalyst was successfully recovered without any loss of heterogeneity and could be reused at least up to the fifth cycle.

Maternal Magnesium Restriction Elevates Glucocorticoid Stress and Inflammation in the Placenta and Fetus of WNIN Rat Dams.

Magnesium plays a major role in many vital functions in the body. We reported earlier that maternal magnesium restriction altered body composition, fat metabolism, and insulin resistance in WNIN rat offspring and was associated with increased glucocorticoid stress in the offspring in their later life. We hypothesize that increased glucocorticoid stress and inflammation which originate in Mg restricted rat dams is transmitted through placenta to the fetus and underlie the metabolic disturbances in the later life of the offspring. Female weanling WNIN rats received ad libitum, a control diet (MgC) or the same with 62% restriction of Mg (MgR) for 3 months, and their plasma magnesium, inflammatory cytokines, and corticosterone were determined (n = 6 per group) before mating. Following mating with control males, placentae, and fetuses were collected on gestational day 15 (GD 15) from MgC and MgR dams (eight dams from each group and three samples from each dam) and used to determine the levels of inflammatory cytokines, corticosterone, and expression of relevant genes. MgR placentae and fetuses had higher (than MgC) levels of corticosterone and proinflammatory cytokines. Expression of Hsd11b1 was increased (sixfold, p < 0.05), while that of Hsd11b2 was decreased (0.4-fold, p < 0.05) in MgR (than MgC) placenta, whereas expression of Hsd11b1was increased (3.4-fold, p < 0.05) in MgR fetus. Chronic dietary magnesium restriction in WNIN female rats increased their levels of corticosterone, leptin, and proinflammatory cytokines which appeared to be transmitted through placenta to the fetus and could thus be associated with increased stress, altered body composition, fat metabolism, and insulin resistance in the later life of the offspring.

Evaluation of Efficacy of 1% Curcuminoids as Local Application in Management of Oral Lichen Planus - Interventional Study.

INTRODUCTION: Oral Lichen Planus (OLP) is a chronic inflammatory mucocutaneous disease affecting the oral cavity which has been associated with severe burning sensation which in turn affects the quality of life of the patients. Currently, the effective treatment of choice remains the use of corticosteroids. The chronicity and recalcitrant nature of the disease demand the long term use of these drugs with ensuing adverse effects. Turmeric and its active ingredient, "curcumin", have been found to have anti-inflammatory, antioxidant and anticancer properties. Hence, this intervention study was done to evaluate the efficacy of curcumin oral gel in the management of oral lichen planus. AIM: The study was conducted to compare the efficacy of 1% curcumin gel with the conventional mid-potent topical corticosteroid, triamcinolone acetonide 0.1% in managing the signs and symptoms of OLP. MATERIALS AND METHODS: An intervention study was conducted on 75 patients diagnosed with OLP divided into three groups, Group 1 (0.1% triamcinolone acetonide oral paste thrice daily in tapering doses), Group 2 (curcumin oral gel thrice daily) and Group 3 (curcumin oral gel six times daily). The patients were analysed for the symptom (burning) and sign (erythema and ulceration) using the numerical rating score and Modified Oral Mucositis Index (MOMI) for a period of three months on a bimonthly basis. The results were analysed using Students t-test and unpaired t-test. RESULTS: Intragroup comparison using paired t-test showed statistically significant reduction in burning sensation (p<0.001) as well as erythema and ulceration (p<0.001) in all the three groups. However, when the three groups were compared Group 1 showed the maximum reduction in burning sensation (77%) and erythema and ulceration (67%). Among the curcuminoid group, increased frequency was found to be more beneficial. CONCLUSION: Curcumin oral gel can bring about clinical improvements in OLP patients; however it cannot be used as a mainstay drug. Instead, it can be used as a maintenance drug after the patient is treated with an initial course of corticosteroids. Further studies with larger sample size and increased drug concentrations may be required.

Characterization and Purification of Neoplastic Cells of Nodular Lymphocyte Predominant Hodgkin Lymphoma from Lymph Nodes by Flow Cytometry and Flow Cytometric Cell Sorting.

We report the flow cytometric (FC) identification and characterization of lymphocyte predominant (LP) cells from tissues involved by nodular lymphocyte predominant Hodgkin lymphoma (NLPHL). First, we immunophenotyped the NLPHL cell line (DEV) confirming a germinal center immunophenotype, lack of expression of CD32 and CD58, and expression of CD54. Nineteen of 26 lymph nodes involved by NLPHL demonstrated a population with an LP immunophenotype (73%), which included expression of germinal center markers (CD75/Bcl-6-positive, CD32-weak/negative without CD10), a B-cell immunophenotype (CD19/CD20/CD40+), IgD and/or IgM expression (67%), and lack of programmed death-ligand 1/ligand 2. The LP cells demonstrated an adhesion macromolecule expression pattern distinct from Hodgkin and Reed-Sternberg cells of classical Hodgkin lymphoma (CHL) (uniform CD50 and variable CD58 for NLPHL; minimal CD50, bright CD58 expression for CHL). A two-tube consensus assay identified LP cells in all seven NLPHL cases examined and only one non-NLPHL case (94 cases evaluated). Finally, FC cell sorting studies confirm that FC-defined populations have an LP cytomorphology. Taken together, these findings demonstrate a two-tube consensus assay can be used to immunophenotype NLHPL with high specificity and sensitivity and rapidly purify LP cells for genetic studies. This study also confirms aneuploidy in LP cells, provides antigens that may be helpful in distinguishing NLPHL from CHL, and suggests that T cells interact less avidly with LP cells than with Hodgkin and Reed-Sternberg cells.

Cutaneous metastasis: An unusual presenting feature of urologic malignancies.

Urological malignancies are well known for their ability to metastasize widely. The incidence of cutaneous metastasis from all urologic malignancies has been reported to be 0.73-1.3% with the primary most commonly being renal cell carcinoma followed by carcinoma bladder, adenocarcinoma prostate, and testicular germ cell tumor in decreasing order of frequency. Metastasis to the skin is unusual and has been predominantly reported as a late manifestation of the disease. We describe two patients with urologic malignancies who had cutaneous metastasis as their initial presenting feature.

A Distributed Network for Intensive Longitudinal Monitoring in Metastatic Triple-Negative Breast Cancer.

Accelerating cancer research is expected to require new types of clinical trials. This report describes the Intensive Trial of OMics in Cancer (ITOMIC) and a participant with triple-negative breast cancer metastatic to bone, who had markedly elevated circulating tumor cells (CTCs) that were monitored 48 times over 9 months. A total of 32 researchers from 14 institutions were engaged in the patient's evaluation; 20 researchers had no prior involvement in patient care and 18 were recruited specifically for this patient. Whole-exome sequencing of 3 bone marrow samples demonstrated a novel ROS1 variant that was estimated to be present in most or all tumor cells. After an initial response to cisplatin, a hypothesis of crizotinib sensitivity was disproven. Leukapheresis followed by partial CTC enrichment allowed for the development of a differential high-throughput drug screen and demonstrated sensitivity to investigational BH3-mimetic inhibitors of BCL-2 that could not be tested in the patient because requests to the pharmaceutical sponsors were denied. The number and size of CTC clusters correlated with clinical status and eventually death. Focusing the expertise of a distributed network of investigators on an intensively monitored patient with cancer can generate high-resolution views of the natural history of cancer and suggest new opportunities for therapy. Optimization requires access to investigational drugs.

Recurrent somatic loss of TNFRSF14 in classical Hodgkin lymphoma.

Investigation of the genetic lesions underlying classical Hodgkin lymphoma (CHL) has been challenging due to the rarity of Hodgkin and Reed-Sternberg (HRS) cells, the pathognomonic neoplastic cells of CHL. In an effort to catalog more comprehensively recurrent copy number alterations occurring during oncogenesis, we investigated somatic alterations involved in CHL using whole-genome sequencing-mediated copy number analysis of purified HRS cells. We performed low-coverage sequencing of small numbers of intact HRS cells and paired non-neoplastic B lymphocytes isolated by flow cytometric cell sorting from 19 primary cases, as well as two commonly used HRS-derived cell lines (KM-H2 and L1236). We found that HRS cells contain strikingly fewer copy number abnormalities than CHL cell lines. A subset of cases displayed nonintegral chromosomal copy number states, suggesting internal heterogeneity within the HRS cell population. Recurrent somatic copy number alterations involving known factors in CHL pathogenesis were identified (REL, the PD-1 pathway, and TNFAIP3). In eight cases (42%) we observed recurrent copy number loss of chr1:2,352,236-4,574,271, a region containing the candidate tumor suppressor TNFRSF14. Using flow cytometry, we demonstrated reduced TNFRSF14 expression in HRS cells from 5 of 22 additional cases (23%) and in two of three CHL cell lines. These studies suggest that TNFRSF14 dysregulation may contribute to the pathobiology of CHL in a subset of cases.