DNA Sequencing of Small Bowel Adenocarcinomas Identifies Targetable Recurrent Mutations in the ERBB2 Signaling Pathway.

PURPOSE: Little is known about the genetic alterations characteristic of small bowel adenocarcinoma (SBA). Our purpose was to identify targetable alterations and develop experimental models of this disease.Experimental Design: Whole-exome sequencing (WES) was completed on 17 SBA patient samples and targeted-exome sequencing (TES) on 27 samples to confirm relevant driver mutations. Two SBA models with ERBB2 kinase activating mutations were tested for sensitivity to anti-ERBB2 agents in vivo and in vitro. Biochemical changes were measured by reverse-phase protein arrays. RESULTS: WES identified somatic mutations in 4 canonical pathways (WNT, ERBB2, STAT3, and chromatin remodeling), which were validated in the TES cohort. Although APC mutations were present in only 23% of samples, additional WNT-related alterations were seen in 12%. ERBB2 mutations and amplifications were present in 23% of samples. Patients with alterations in the ERBB2 signaling cascade (64%) demonstrated worse clinical outcomes (median survival 70.3 months vs. 109 months; log-rank HR = 2.4, P = 0.03). Two ERBB2-mutated (V842I and Y803H) cell lines were generated from SBA patient samples. Both demonstrated high sensitivity to ERBB2 inhibitor dacomitinib (IC50 < 2.5 nmol/L). In xenografts derived from these samples, treatment with dacomitinib reduced tumor growth by 39% and 59%, respectively, whereas it had no effect in an SBA wild-type ERBB2 model. CONCLUSIONS: The in vitro and in vivo models of SBA developed here provide a valuable resource for understanding targetable mutations in this disease. Our findings support clinical efforts to target activating ERBB2 mutations in patients with SBA that harbor these alterations.

Activation of persons living with HIV for treatment, the great study.

BACKGROUND: Patient empowerment represents a potent tool for addressing racial, ethnic and socioeconomic disparities in health care, particularly for chronic conditions such as HIV infection that require active patient engagement. This multimodal intervention, developed in concert with HIV patients and clinicians, aims to provide HIV patients with the knowledge, skills, attitudes and tools to become more activated patients. METHODS/DESIGN: Randomized controlled trial of a multimodal intervention designed to activate persons living with HIV. The intervention includes four components: 1) use of a web-enabled hand-held device (Apple iPod Touch) loaded with a Personal Health Record (ePHR) customized for HIV patients; 2) six 90-minute group-based training sessions in use of the device, internet and the ePHR; 3) a pre-visit coaching session; and 4) clinician education regarding how they can support activated patients. Outcome measures include pre- post changes in patient activation measure score (primary outcome), eHealth literacy, patient involvement in decision-making and care, medication adherence, preventive care, and HIV Viral Load. DISCUSSION: We hypothesize that participants receiving the intervention will show greater improvement in empowerment and the intervention will reduce disparities in study outcomes. Disparities in these measures will be smaller than those in the usual care group. Findings have implications for activating persons living with HIV and for other marginalized groups living with chronic illness. TRIAL REGISTRATION: ClinicalTrials.gov Identifier: NCT02165735, 6/13/2014.

Management of the Potential Organ Donor in the ICU: Society of Critical Care Medicine/American College of Chest Physicians/Association of Organ Procurement Organizations Consensus Statement.

This document was developed through the collaborative efforts of the Society of Critical Care Medicine, the American College of Chest Physicians, and the Association of Organ Procurement Organizations. Under the auspices of these societies, a multidisciplinary, multi-institutional task force was convened, incorporating expertise in critical care medicine, organ donor management, and transplantation. Members of the task force were divided into 13 subcommittees, each focused on one of the following general or organ-specific areas: death determination using neurologic criteria, donation after circulatory death determination, authorization process, general contraindications to donation, hemodynamic management, endocrine dysfunction and hormone replacement therapy, pediatric donor management, cardiac donation, lung donation, liver donation, kidney donation, small bowel donation, and pancreas donation. Subcommittees were charged with generating a series of management-related questions related to their topic. For each question, subcommittees provided a summary of relevant literature and specific recommendations. The specific recommendations were approved by all members of the task force and then assembled into a complete document. Because the available literature was overwhelmingly comprised of observational studies and case series, representing low-quality evidence, a decision was made that the document would assume the form of a consensus statement rather than a formally graded guideline. The goal of this document is to provide critical care practitioners with essential information and practical recommendations related to management of the potential organ donor, based on the available literature and expert consensus.

Preliminary whole-exome sequencing reveals mutations that imply common tumorigenicity pathways in multiple endocrine neoplasia type 1 patients.

BACKGROUND: Whole-exome sequencing studies have not established definitive somatic mutation patterns among patients with sporadic hyperparathyroidism (HPT). No sequencing has evaluated multiple endocrine neoplasia type 1 (MEN1)-related HPT. We sought to perform whole-exome sequencing in HPT patients to identify somatic mutations and associated biological pathways and tumorigenic networks. METHODS: Whole-exome sequencing was performed on blood and tissue from HPT patients (MEN1 and sporadic) and somatic single nucleotide variants (SNVs) were identified. Stop-gain and stop-loss SNVs were analyzed with Ingenuity Pathways Analysis (IPA). Loss of heterozygosity (LOH) was also assessed. RESULTS: Sequencing was performed on 4 MEN1 and 10 sporadic cases. Eighteen stop-gain/stop-loss SNV mutations were identified in 3 MEN1 patients. One complex network was identified on IPA: Cellular function and maintenance, tumor morphology, and cardiovascular disease (IPA score = 49). A nonsynonymous SNV of TP53 (lysine-to-glutamic acid change at codon 81) identified in a MEN1 patient was suggested to be a driver mutation (Cancer-specific High-throughput Annotation of Somatic Mutations; P = .002). All MEN1 and 3/10 sporadic specimens demonstrated LOH of chromosome 11. CONCLUSION: Whole-exome sequencing revealed somatic mutations in MEN1 associated with a single tumorigenic network, whereas sporadic pathogenesis seemed to be more diverse. A somatic TP53 mutation was also identified. LOH of chromosome 11 was seen in all MEN1 and 3 of 10 sporadic patients.

Analysis of autoantibodies to 3-hydroxy-3-methylglutaryl-coenzyme A reductase using different technologies.

Diagnostic tests are needed to aid in the diagnosis of necrotizing myopathies associated with statin use. This study aimed to compare different technologies for the detection of anti-HMGCR antibodies and analyze the clinical phenotype and autoantibody profile of the patients. Twenty samples from myositis patients positive for anti-HMGCR antibodies using a research addressable laser bead assay and 20 negative controls were tested for autoantibodies to HMGCR: QUANTA Lite HMGCR ELISA and QUANTA Flash HMGCR CIA. All patients were also tested for antibodies to extractable nuclear antigens and myositis related antibodies. To verify the specificity of the ELISA, 824 controls were tested. All three assays showed qualitative agreements of 100% and levels of anti-HMGCR antibodies showed significant correlation: Spearman's rho > 0.8. The mean age of the anti-HMGCR antibody positive patients was 54.4 years, 16/20 were females, and 18/20 had necrotizing myopathy (two patients were not diagnosed). Nine out of 20 anti-HMGCR positive patients were on statin. All patients with anti-HMGCR antibodies were negative for all other autoantibodies tested. Testing various controls showed high specificity (99.3%). Anti-HMGCR antibodies are not always associated with the use of statin and appear to be the exclusive autoantibody specificity in patients with statin associated myopathies.

Bridging the digital divide in HIV care: a pilot study of an iPod personal health record.

BACKGROUND: Persons living with HIV (PLWH) need practical tools to self-manage their condition. METHODS: We conducted a proof-of-concept study among PLWH to assess whether patients could learn to use a personal health record (PHR) on a hand-held device (iPod Touch) to manage their condition. We began individual trainings and later adapted this to group training. We assessed usability, acceptability and also effects on self-efficacy for treatment adherence using the HIV Treatment Adherence Self-Efficacy Scale (HIV-ASES). RESULTS: Nine PLWH participated in the individual training and 29 participated in the group sessions. The participants were largely middle aged, low-income and of racial/ethnic minorities. The sessions were well attended and participants fully engaged in tasks and shared learning. Most participants stated they intended to use the PHR and reported improved self-efficacy in treatment adherence (P = .05) particularly on the integration of treatment adherence into one's routine (P < .02). CONCLUSIONS: Training PLWH in use of a handheld PHR shows promise.

Efficacy of renal artery angioplasty and stenting in a solitary functioning kidney.

OBJECTIVE: The aim of this study was to share our experience of percutaneous renal artery angioplasty and stenting (PTRAS) in our patients with a solitary functioning kidney over a 10-year period. METHODS: The procedures were performed on 75 patients from 1995 to 2004. Data were collected retrospectively from case notes of patients. The definition for solitary functioning kidney was a contralateral kidney size of <8 cm, complete occlusion of contralateral renal artery or previous nephrectomy. Serum creatinine was considered improved or worse if the deviation from the baseline value was >20%. RESULTS: For the purpose of halting renal deterioration (n=47), there were improvement and stabilization in 21% and 55% at 3 months and 28% and 28% at 12 months. Systolic blood pressure (n=27) improvement and stabilization were achieved in 33% and 56% both at 3 and 12 months. Diastolic blood pressure (n=27) improvement and stabilization were 22% and 70% at 3 months and 33% and 48% at 12 months. Five out of seven patients with acute renal failure (serum creatinine>500 micromol/l and requiring haemodialysis) pre-procedure were dialysis-free at 12 months. Complications occurred in 19 (25%) patients and these included bleeding (n=16), pseudoaneurysm (n=3), renal artery dissection (n=2) and cholesterol embolization (n=1). CONCLUSION: PTRAS in a solitary functioning kidney produced clinical benefits in the majority of patients with resistant hypertension and renal function deterioration.

Structure of nucleotide-binding domain 1 of the cystic fibrosis transmembrane conductance regulator.

Cystic fibrosis transmembrane conductance regulator (CFTR) is an ATP-binding cassette (ABC) transporter that functions as a chloride channel. Nucleotide-binding domain 1 (NBD1), one of two ABC domains in CFTR, also contains sites for the predominant CF-causing mutation and, potentially, for regulatory phosphorylation. We have determined crystal structures for mouse NBD1 in unliganded, ADP- and ATP-bound states, with and without phosphorylation. This NBD1 differs from typical ABC domains in having added regulatory segments, a foreshortened subdomain interconnection, and an unusual nucleotide conformation. Moreover, isolated NBD1 has undetectable ATPase activity and its structure is essentially the same independent of ligand state. Phe508, which is commonly deleted in CF, is exposed at a putative NBD1-transmembrane interface. Our results are consistent with a CFTR mechanism, whereby channel gating occurs through ATP binding in an NBD1-NBD2 nucleotide sandwich that forms upon displacement of NBD1 regulatory segments.

Glucose sensing in transdermal body fluid collected under continuous vacuum pressure via micropores in the stratum corneum.

Application of continuous vacuum pressure on skin perforated with tiny micropores created by a focused beam from a low-cost laser system can result in access to a clear, transdermal body fluid (TDF) for the continuous measurement of glucose in vivo. Two clinical studies were performed to assess the feasibility of this approach. In the first study, 56 diabetic subjects were porated on either the arm or abdomen, and glucose was measured in their TDF using a custom assay system contained in a patch that was affixed to the skin above the poration site. The continuous readings of glucose in TDF were compared with fingerstick blood measured every half-hour over a 2-day period, resulting in 1,167 paired data points that yielded a correlation of 0.8745 with 97.75% of the readings in the Clarke Error Grid A and B zones. In a second study, 187 diabetic and 65 nondiabetic subjects had glucose measurements from their TDF made using a commercially available glucose strip and meter. A total of 4,059 data pairs (discrete TDF and capillary blood) were collected over a 2-day period, resulting in a correlation of 0.946 with 99% of the readings in the Clarke Error Grid A and B zones. These studies indicate that TDF drawn through micropores in the stratum corneum of the skin potentially can provide a lesser invasive and continuous method of measuring glucose in diabetic individuals.

Atherosclerotic renal arterial stenosis: clinical outcomes of stent placement for hypertension and renal failure.

PURPOSE: To assess technical success rates and long-term clinical outcomes of primary renal arterial stent placement in atherosclerotic renal arterial stenosis (RAS). MATERIALS AND METHODS: Primary stent placement was performed in 100 consecutive patients with atherosclerotic RAS. Indications for treatment were resistant hypertension (n = 25), impaired renal function, (n = 50), and both (n = 25). Immediate technical results were evaluated with angiography. Clinical outcomes were assessed with serial systolic and diastolic blood pressure and serum creatinine values obtained from retrospective review of case notes. Results obtained every 6 months after the procedure were compared with those obtained at the time of the procedure with the paired t test. Radiologic reports were evaluated for immediate and case notes for delayed complications. RESULTS: Technical success was achieved in 120 (95.2%) of 126 RAS in 95 patients. Mean follow-up was 25 months (median, 24 months; range, 1-66 months). Resistant hypertension was cured in two (4.2%) of 48 patients, had improved in 38 (79.1%), and had failed to respond to treatment in eight (16.7%). Mean systolic and diastolic blood pressures were significantly lower at 6, 12, 18, 24, and 30 months (P <.01) than before the procedure. Among 65 patients treated for renal impairment, renal function improved in 20 (30.8%), stabilized in 25 (41.7%), and continued to deteriorate in 20 (30.8%). The mean serum creatinine level did not show significant change with time for this group. In the improved subgroup, it was significantly higher at 6, 12, 18, 24, 36, and 42 months (P <.05) than prior to the procedure. Procedure-related complications occurred in 18 (18%) cases: Ten were minor and self-limiting and eight were major and included two procedure-related deaths. CONCLUSION: In atherosclerotic RAS, primary stent deployment has a high technical success rate, producing clinical benefits in the majority of patients when performed for resistant hypertension and recovery of renal function.