Hepatitis C virus infects and perturbs liver stem cells.

Hepatitis C virus (HCV) is the leading cause of death from liver disease. How HCV infection causes lasting liver damage and increases cancer risk remains unclear. Here, we identify bipotent liver stem cells as novel targets for HCV infection, and their erroneous differentiation as the potential cause of impaired liver regeneration and cancer development. We show 3D organoids generated from liver stem cells from actively HCV-infected individuals carry replicating virus and maintain low-grade infection over months. Organoids can be infected with a primary HCV isolate. Virus-inclusive single-cell RNA sequencing uncovered transcriptional reprogramming in HCV+ cells supporting hepatocytic differentiation, cancer stem cell development, and viral replication while stem cell proliferation and interferon signaling are disrupted. Our data add a new pathogenesis mechanism-infection of liver stem cells-to the biology of HCV infection that may explain progressive liver damage and enhanced cancer risk through an altered stem cell state.ImportanceThe hepatitis C virus (HCV) causes liver disease, affecting millions. Even though we have effective antivirals that cure HCV, they cannot stop terminal liver disease. We used an adult stem cell-derived liver organoid system to understand how HCV infection leads to the progression of terminal liver disease. Here, we show that HCV maintains low-grade infections in liver organoids for the first time. HCV infection in liver organoids leads to transcriptional reprogramming causing cancer cell development and altered immune response. Our finding shows how HCV infection in liver organoids mimics HCV infection and patient pathogenesis. These results reveal that HCV infection in liver organoids contributes to liver disease progression.

Modelling T-cell immunity against hepatitis C virus with liver organoids in a microfluidic coculture system.

Hepatitis C virus (HCV) remains a global public health challenge with an estimated 71 million people chronically infected, with surges in new cases and no effective vaccine. New methods are needed to study the human immune response to HCV since in vivo animal models are limited and in vitro cancer cell models often show dysregulated immune and proliferative responses. Here, we developed a CD8+ T cell and adult stem cell liver organoid system using a microfluidic chip to coculture 3D human liver organoids embedded in extracellular matrix with HLA-matched primary human T cells in suspension. We then employed automated phase contrast and immunofluorescence imaging to monitor T cell invasion and morphological changes in the liver organoids. This microfluidic coculture system supports targeted killing of liver organoids when pulsed with a peptide specific for HCV non-structural protein 3 (NS3) (KLVALGINAV) in the presence of patient-derived CD8+ T cells specific for KLVALGINAV. This demonstrates the novel potential of the coculture system to molecularly study adaptive immune responses to HCV in an in vitro setting using primary human cells.

Lisdexamfetamine Pharmacokinetic Comparison Between Patients Who Underwent Roux-en-Y Gastric Bypass and Nonsurgical Controls.

INTRODUCTION/PURPOSE: The objective of this research was to characterize the impact of Roux-en-Y gastric bypass (RYGB) on the pharmacokinetic properties of the pro-drug lisdexamfetamine and its active metabolite, d-amphetamine. MATERIALS AND METHODS: A case-control design was used where patients who had undergone RYGB 9-24 months prior were matched on sex, age, and body mass index (BMI) to nonsurgical controls who had no history of weight loss surgery. Each participant received a single 50 mg dose of lisdexamfetamine, and plasma samples were collected over a 24-h period following dosing. Noncompartmental analyses were used to compare pharmacokinetic measures between groups. RESULTS: There were no significant differences between the RYGB (n = 10) and NSC groups (n = 10) on sex (70% female), age (40.9 ± 9.6 vs. 41.3 ± 8.9 years), BMI (30.3 ± 5.2 vs. 31 ± 5.9 kg/m2), or ethnicity (100% vs. 80% White). The pharmacokinetic parameters between the RYGB and NCS groups were found to be equivalent for lisdexamfetamine and d-amphetamine, including maximum plasma concentration (Cmax), time to maximum plasma concentration (Tmax), and area under the plasma concentration-time curve (AUC(0-∞)). CONCLUSION: These data suggest that there is no need to routinely adjust lisdexamfetamine dosing following RYGB. However, given the potential for inter-individual differences, patients who undergo RYGB should be clinically monitored and individualized dosing strategies should be considered for concerns surrounding efficacy or toxicity.

The Evaluation of Screening Questionnaires for Obstructive Sleep Apnea to Identify High-Risk Obese Patients Undergoing Bariatric Surgery.

BACKGROUND: Obstructive sleep apnea (OSA) is extremely common among bariatric surgical candidates. Identifying those at risk for moderate to severe OSA is challenging. Testing all bariatric surgical candidates with a level 1 polysomnographic study is expensive and resource intensive. The aim of this study is to evaluate three standardized screening questionnaires that are utilized to identify high-risk patients for OSA undergoing bariatric surgery. METHODS: A retrospective review of data collected prospectively was undertaken on bariatric surgical patients who have not had a preexisting diagnosis of OSA. Each patient was subjected to the STOP BANG and Berlin Questionnaires as well as the Epworth Sleepiness Scale (ESS), after which a level 1 polysomnogram was undertaken. Nonparametric receiver operating characteristic analyses were used to evaluate the relationship between questionnaire scores and OSA as determined by a formal sleep lab study. RESULTS: There were 266 patients subjected to a standard overnight polysomnogram and screening questionnaires. Area under the curve (AUC) values for analyses including the entire sample were significantly (p < .05) greater than chance (i.e., AUC = .50) for all questionnaire scores except the ESS for both severe OSA (AUC range = .584-.631) and moderate/severe OSA (AUC range = .589-.660), although the magnitude of the AUC values was quite modest. Sensitivity and specificity values from the current study are substantially lower than those previously reported in the literature. CONCLUSIONS: Neither the STOP BANG nor Berlin questionnaires appear to be effective tools for detecting moderate- or high-risk patients for OSA undergoing bariatric surgery.

Examining the role of self-discrepancy and self-directed style in bulimia nervosa.

Two of the primary components within Integrative Cognitive Affective Therapy (ICAT) are self-discrepancy and self-directed style. Self-discrepancy includes both actual:ideal (discrepancy between oneself and who one wishes they were) and actual:ought (discrepancy between oneself and who one believes they ought to be). Self-directed style in ICAT refers to a variety of behaviors emitted by a person toward the self including self-blaming and self-affirming. This study explored main effects and interactions between self-discrepancy and self-directed style in relation to global eating disorder (ED) psychopathology, depressive symptoms, and anxiety. Eighty treatment-seeking adults from the Midwest with BN or subthreshold BN completed interviews and self-report measures. Self-affirm and self-blame were associated with ED psychopathology, depressive symptoms, and anxiety. Actual:ideal discrepancy was related to anxiety and actual:ought discrepancy was related to anxiety and depressive symptoms. Interactions were found between self-affirm and actual:ought discrepancy as well as self-blame and actual:ought discrepancy for depressive symptoms. High actual:ought was related to increased depressive symptoms regardless of levels of self-affirm or self-blame. Effect sizes for models were medium-to-large with anxiety models demonstrating the largest effects. This study provides further evidence supporting the ICAT model and treatment, which targets self-discrepancies, self-directed styles, and related emotional states.

Comparison of Warfarin Dosages and International Normalized Ratios Before and After Roux-en-Y Gastric Bypass Surgery.

STUDY OBJECTIVE: As data are limited for dosing warfarin in patients who have undergone bariatric surgery, our objective was to gather data on warfarin pharmacotherapy to aid in the prescribing practices and dosage adjustment of warfarin after Roux-en-Y gastric bypass (RYGB) surgery. DESIGN: Retrospective medical record review. SETTING: Anticoagulation clinic. PATIENTS: Twelve patients who underwent RYGB surgery between 2009 and 2011. MEASUREMENTS AND MAIN RESULTS: Data were collected on patient demographics, therapeutic indications for warfarin, weekly warfarin doses, and international normalized ratio (INR) values before and after RYGB surgery. To examine whether the relationship between the warfarin dose and therapeutic effect (i.e., INR) of warfarin changed from before to after surgery, we generated a ratio between each INR measurement and the weekly warfarin dose that immediately preceded it (warfarin dose/INR ratio). Mean weekly warfarin dose after RYGB surgery decreased significantly relative to presurgery dose (estimated marginal mean ± SE presurgery dose 37.08 ± 3.31 mg vs postsurgery dose 28.08 ± 3.27 mg; [F(205.63) = 13.23, p<0.001]), whereas INRs increased significantly after surgery from presurgery levels (estimated marginal mean ± SE presurgery INR 2.36 ± 0.15 vs postsurgery INR 2.94 ± 0.14; [F(213.76) = 6.55, p<0.05]). The ratio of warfarin dose/INR from presurgery (estimated marginal mean ± SE 17.64 ± 1.67) to postsurgery (estimated marginal mean ± SE 10.94 ± 1.64) was also significantly reduced [F(206.47) = 20.61, p<0.001], suggesting that a lower warfarin dose was required to produce the desired INR after surgery compared with before surgery. CONCLUSION: Warfarin dosages tended to decrease after RYGB surgery, in agreement with previously published literature. These findings suggest the need for increased INR monitoring and patient education for patients taking warfarin after RYGB surgery.

Survey on laparoscopic sleeve gastrectomy (LSG) at the Fourth International Consensus Summit on Sleeve Gastrectomy.

BACKGROUND: LSG has been increasingly performed. Long-term follow-up is necessary. METHODS: During the Fourth International Consensus Summit on LSG in New York Dec. 2012, an online questionnaire (SurveyMonkey®) was filled out by 130 surgeons experienced in LSG. The survey was submitted directly to the statisticians. RESULTS: The 130 surgeons performed 354.9 ± SD 453 LSGs/surgeon (median 175), for a total of 46,133 LSGs. The LSGs had been performed over 4.9 ± 2.7 year (range 1-10). Of the 46,133 LSGs, 0.2 ± 1.0 % (median 0, range 0-10 %) were converted to an open operation. LSG was intended as the sole operation in 93.1 ± 14.8 %; in 3.0 ± 6.3 %, a second stage became necessary. Of the 130 surgeons, 40 (32 %) use a 36F bougie, which was most common (range 32-50F). Staple-line is reinforced by 79 %; of these, 57 % use a buttress and 43 % over-sew. Mean %EWL at year 1 was 59.3 %; year 2, 59.0 %; year 3, 54.7 %; year 4, 52.3 %; year 5, 52.4 %; and year 6, 50.6 %. If a second-stage operation becomes necessary, preference was: RYGB 46 %, duodenal switch 24 %, re-sleeve 20 %, single-anastomosis duodenoileal bypass 3 %, sleeve plication 3 %, minigastric bypass 3 %, non-adjustable band 2 %, and side-to-side jejunoileal anastomosis 1 %. Complications were: high leak 1.1 %, hemorrhage 1.8 %, and stenosis at lower sleeve 0.9 %. Postoperative gastroesophageal reflux occurred in 7.9 ± 8.2 % but was variable (0-30 %). Mortality was 0.33 ± 1.6 %, which translates to ≈ 152 deaths. Eighty-nine percent order multivitamins (including vitamin D, calcium, and iron) and 72 % order B12. A PPI is ordered by 29 % for 1 month, 29 % for 3 months, and others for 1-12 months depending on the case. CONCLUSIONS: LSG was relatively safe. Further long-term surveillance is necessary.

Third International Summit: Current status of sleeve gastrectomy.

BACKGROUND: Laparoscopic sleeve gastrectomy (LSG) has been performed for morbid obesity in the past 10 years. LSG was originally intended as a first-stage procedure in high-risk patients but has become a stand-alone operation for many bariatric surgeons. Ongoing review is necessary regarding the durability of the weight loss, complications, and need for second-stage operations. METHODS: The first International Summit for LSG was held in October 2007, the second in March 2009, and this third in December 2010. There were presentations by experts, and, to provide a consensus, a questionnaire was completed by 88 attendees who had >1 year (mean 3.6 ± 1.5, range 1-8) of experience with LSG. RESULTS: The results of the questionnaire were based on 19,605 LSGs performed within 3.6 ± 1.5 years (228.8 ± 275.0 LSGs/surgeon). LSG had been intended as the sole operation in 86.4% of the cases; in these, a second-second stage became necessary in 2.2%. LSG was completed laparoscopically in 99.7% of the cases. The mean percentage of excess weight loss at 1, 2, 3, 4, and 5 years was 62.7%, 64.7%, 64.0%, 57.3%, and 60.0%, respectively. The bougie size was 28-60F (mean 36F, 70% blunt tip). Resection began 1.5-7.0 cm (mean 4.8) proximal to the pylorus. Of the surgeons, 67.1% reinforced the staple line, 57% with buttress material and 43% with oversewing. The respondents excised an estimated 92.9% ± 8.0% (median 95.0%) of fundus (i.e., a tiny portion is maintained lateral to the angle of His). A drain is left by 57.6%, usually closed suction. High leaks occurred in 1.3% of cases (range 0-10%); lower leaks occurred in .5%. Intraluminal bleeding occurred in 2.0% of cases. The mortality rate was .1% ± .3%. CONCLUSION: According to the questionnaire, presentations, and debates, the weight loss and improvement in diabetes appear to be better than with laparoscopic adjustable gastric banding and on par with Roux-en-Y gastric bypass. High leaks are infrequent but problematic.

Evidence-based mental health interventions for traumatized youth: a statewide dissemination project.

Psychological trauma in childhood has been shown to increase a variety of psychological disturbances and psychiatric disorders. Although evidence-based treatments for children who have been traumatized exist, they are infrequently used by clinicians treating children. The present paper describes the creation of the Treatment Collaborative for Traumatized Youth (TCTY) which is a statewide partnership in North Dakota designed to disseminate efficacious treatments for traumatized children and monitor outcomes across a broad, rural, geographic expanse. The paper reviews the dissemination strategy developed by the TCTY, reports outcomes regarding both clinicians and child participants, and highlights problems identified in the project and solutions that were generated.

A prevalence study and description of alli use by patients with eating disorders.

OBJECTIVE: This study examined the frequency and characteristics of alli use among patients in eating disorder treatment facilities. METHOD: Patients from five treatment centers completed the Survey of Eating and Related Behaviors. Diagnoses were determined from survey responses. RESULTS: Of 417 survey respondents, 26 (6.2%) reported a history of alli use. Of those, 15 (57.7%) met criteria for an eating disorder, including one of 29 patients (3.4%) with anorexia nervosa binge-purge subtype, six of 66 patients (9.1%) with full or subthreshold bulimia nervosa, four of 49 (8.2%) with binge eating disorder, one of six (16.7%) with purging disorder, and three of 80 (3.8%) with an eating disorder not otherwise specified. DISCUSSION: The results of this survey suggest that patients with eating disorders use alli, albeit relatively uncommonly. Therefore, it is worthwhile for clinicians to inquire about alli use when evaluating or treating these patients in any clinical setting.

The Second International Consensus Summit for Sleeve Gastrectomy, March 19-21, 2009.

BACKGROUND: Sleeve gastrectomy (SG) is a rapid and comparatively simple bariatric operation, which thus far shows good resolution of co-morbidities and good weight loss. The potential peri-operative complications must be recognized and treated promptly. Like other bariatric operations, there are variations in technique. Laparoscopic SG was initially performed for high-risk patients to increase the safety of a second operation. However, indications for SG have been increasing. Interaction among those performing this procedure is necessary, and the Second International Consensus Summit for SG (ICSSG) was held to evaluate techniques and results. METHODS: A questionnaire was filled out by attendees at the Second ICSSG, held March 19-22, 2009, in Miami Beach, and rapid responses were recorded during the consensus part. RESULTS: Findings are based on 106 questionnaires representing a total of 14,776 SGs. In 86.3%, SG was intended as the sole operation. A total of 81.9% of the surgeons reported no conversions from a laparoscopic to an open SG. Mean +/- SD percent excess weight loss was as follows: 1 year, 60.7 +/- 15.6; 2 years, 64.7 +/- 12.9; 3 years, 61.7 +/- 11.4; 4 years 64.6 +/- 10.5; >4 years, 48.5 +/- 8.7. Bougie size was 35.6F +/- 4.9F (median 34.0F, range 16F-60F). The dissection commenced 5.0 +/- 1.4 cm (median 5.0 cm, range 1-10 cm) proximal to the pylorus. Staple-line was reinforced by 65.1% of the responders; of these, 50.9% over-sew, 42.1% buttress, and 7% do both. Estimated percent of fundus removed was 95.8 +/- 12%; many expressed caution to avoid involving the esophagus. Post-operatively, a high leak occurred in 1.5%, a lower leak in 0.5%, hemorrhage in 1.1%, splenic injury in 0.1%, and later stenosis in 0.9%. Post-operative gastroesophageal reflux ( approximately 3 mo) was reported in 6.5% (range 0-83%). Mortality was 0.2 +/- 0.9% (total 30 deaths in 14,776 patients). During the consensus part, the audience responded that there was enough evidence published to support the use of SG as a primary procedure to treat morbid obesity and indicated that it is on par with adjustable gastric banding and Roux-en-Y gastric bypass, with a yes vote at 77%. CONCLUSION: SG for morbid obesity is very promising as a primary operation.

The effect of age on weight-related quality of life in overweight and obese individuals.

The objective of the current study was to investigate the association between age and weight-related quality of life in a broad range of overweight/obese individuals. Participants included 9,991 overweight and obese adults from a cross sectional database (mean age = 44.9, mean BMI = 38.3, 75.3% women, 73% white). Participants completed the Impact of Weight on Quality of Life-Lite (IWQOL-Lite), a measure of weight-related quality of life. For the total sample, weight-related quality of life was more impaired with increasing age for physical function, sexual life, and work. However, increasing age was associated with less impairment for self-esteem and public distress. On the sexual life domain there was an interaction between age and gender. Men showed a steady decline in sexual life with increasing age, whereas women showed reduced scores on sexual life in all age groups beyond age 18-24.9. Of note, women's scores on all IWQOL-Lite domains were significantly lower (more impaired) than men's. Thus, there are both positive as well as negative consequences of increasing age with respect to the impact of weight on quality of life in overweight and obese persons.

T cell responses to human endogenous retroviruses in HIV-1 infection.

Human endogenous retroviruses (HERVs) are remnants of ancient infectious agents that have integrated into the human genome. Under normal circumstances, HERVs are functionally defective or controlled by host factors. In HIV-1-infected individuals, intracellular defense mechanisms are compromised. We hypothesized that HIV-1 infection would remove or alter controls on HERV activity. Expression of HERV could potentially stimulate a T cell response to HERV antigens, and in regions of HIV-1/HERV similarity, these T cells could be cross-reactive. We determined that the levels of HERV production in HIV-1-positive individuals exceed those of HIV-1-negative controls. To investigate the impact of HERV activity on specific immunity, we examined T cell responses to HERV peptides in 29 HIV-1-positive and 13 HIV-1-negative study participants. We report T cell responses to peptides derived from regions of HERV detected by ELISPOT analysis in the HIV-1-positive study participants. We show an inverse correlation between anti-HERV T cell responses and HIV-1 plasma viral load. In HIV-1-positive individuals, we demonstrate that HERV-specific T cells are capable of killing cells presenting their cognate peptide. These data indicate that HIV-1 infection leads to HERV expression and stimulation of a HERV-specific CD8+ T cell response. HERV-specific CD8+ T cells have characteristics consistent with an important role in the response to HIV-1 infection: a phenotype similar to that of T cells responding to an effectively controlled virus (cytomegalovirus), an inverse correlation with HIV-1 plasma viral load, and the ability to lyse cells presenting their target peptide. These characteristics suggest that elicitation of anti-HERV-specific immune responses is a novel approach to immunotherapeutic vaccination. As endogenous retroviral sequences are fixed in the human genome, they provide a stable target, and HERV-specific T cells could recognize a cell infected by any HIV-1 viral variant. HERV-specific immunity is an important new avenue for investigation in HIV-1 pathogenesis and vaccine design.