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Purpose: Men receiving androgen deprivation therapy for prostate cancer have low knowledge of osteoporosis (OP) and engage in few healthy bone behaviors (HBBs). A multicomponent intervention was piloted in this population. Changes in OP knowledge, self-efficacy, health beliefs, and engagement in HBBs were evaluated., Methods: A pre-post pilot study was performed in a convenience sample of men recruited from the Princess Margaret Cancer Centre. Men were sent personalized letters explaining their dual x-ray absorptiometry (DXA) results and fracture risk assessment with an OP-related education booklet. Participants completed questionnaires assessing OP knowledge, self-efficacy, health beliefs, and current engagement in HBBs at baseline (T1) and 3 months post-intervention (T2). Paired t tests and McNemar's test were used to assess changes in outcomes., Results: A total of 148 men completed the study. There was an increase in OP knowledge (9.7 ± 4.3 to 11.4 ± 3.3, p < 0.0001) and feelings of susceptibility (16.5 ± 4.3 to 17.4 ± 4.7, p = 0.015), but a decrease in total self-efficacy (86.3 ± 22.9 to 81.0 ± 27.6, p = 0.007) from baseline to post-intervention. Men made appropriate changes in their overall daily calcium intake (p ≤ 0.001), and there was uptake of vitamin D supplementation from 44 % (n = 65) to 68 % (n = 99) (p < 0.0001). Men with bone loss (osteopenia or OP) had a greater change in susceptibility (1.9 ± 4.3 vs. -0.22 ± 4.2, p = 0.005) compared to men with normal bone density., Conclusions: Our results provide preliminary evidence that a multicomponent intervention such as the one described can lead to increased knowledge and feelings of susceptibility regarding OP and can enhance uptake of some HBBs.

The thrombospondins (TSPs) are a family of matricellular proteins that regulate cellular phenotype through interactions with a myriad of other proteins and proteoglycans. We have identified a novel interaction of the members of the TSP gene family with stromal interaction molecule 1 (STIM1). This association is robust since it is preserved in Triton X-100, can be detected with multiple anti-TSP-1 and anti-STIM1 antibodies, and is detected in a wide range of cell types. We have also found that STIM1 co-immunoprecipitates with TSP-4 and cartilage oligomeric matrix protein (COMP), and that a recombinant version of the N-terminal domain of STIM1 binds to the signature domain of TSP-1 and COMP. The association of the TSPs with STIM1 is observed in both the presence and absence of calcium indicating that the calcium-dependent conformation of the signature domain of TSPs is not required for binding. Thus, this interaction could occur in the ER under conditions of normal or low calcium concentration. Furthermore, we observed that the expression of COMP in HEK 293 cells decreases STIM1-mediated calcium release activated calcium (CRAC) channel currents and increases arachidonic acid calcium (ARC) channel currents. These data indicate that the TSPs regulate STIM1 function and participate in the reciprocal regulation of two channels that mediate calcium entry into the cell., (Copyright © 2014 The Authors. Published by Elsevier B.V. All rights reserved.)

Acute gastrointestinal (GI) bleeding is a common cause of hospitalization, resulting in about 400,000 hospital admissions annually, with a mortality rate of 5-10%. It is estimated that 5% of acute GI bleedings are of obscure origin with a normal esophagogastroduodenoscopy and ileocolonoscopy. Capsule endoscopy is the state-of-the-art procedure for inspection of the entire small bowel with a high sensitivity for the detection of causes of bleeding. In recent years, many studies have addressed the sensitivity and outcome of capsule-endoscopy procedures in patients with acute GI bleeding. This review looks at the role of capsule endoscopy in the evaluation of patients with acute GI bleeding from either the upper GI tract or small bowel.

Child maltreatment is a leading cause of pediatric morbidity and mortality, described as one of the greatest threats facing the health, welfare, and social well-being of children in the United States (U.S. Department of Health and Human Services, 2012). Despite mandatory reporting laws, the poignant reality is that this public health problem is significantly underrecognized and underreported by heath care providers. The purpose of this project was to implement a series of strategies in a pediatric emergency department to identify children who are at risk of maltreatment and initiate interventions to ensure their safety and protection before a devastating outcome occurs. The results of this project support the implementation of nursing education, a screening program for risk of child maltreatment, and collaboration with interdisciplinary stakeholders to achieve best practice in emergency medicine.

Successful translation of findings derived from preclinical studies into effective therapies is critical in biomedical research. Lack of robustness and reproducibility of the preclinical data, due to insufficient number of repeats, inadequate cell-based and mouse models contribute to the poor success rate. Antibodies are widely used in preclinical research, notably to determine the expression of potential therapeutic targets in tissues of interest, including tumors, but also to identify disease and/or treatment response biomarkers. We sought to determine whether the current antibody characterization standards in preclinical research are sufficient to ensure reliability of the data found in peer-reviewed publications. To address this issue, we used detection of the protein c-FLIP, a major factor of resistance to apoptosis, as a proof of concept. Accurate detection of endogenous c-FLIP levels in the preclinical settings is imperative since it is considered as a potential theranostic biomarker. Several sources of c-FLIP antibodies validated by their manufacturer and recommended for western blotting were therefore rigorously tested. We found a wide divergence in immune recognition properties. While these antibodies have been used in many publications, our results show that several of them failed to detect endogenous c-FLIP protein by Western blotting. Our results suggest that antibody validation standards are inadequate, and that systematic use of genetic knockdowns and/or knockouts to establish proof of specificity is critical, even for antibodies previously used in the scientific literature. Because antibodies are fundamental tools in both preclinical and clinical research, ensuring their specificity is crucial.

Objectives: To describe in patients with prostate cancer, receiving androgen-deprivation therapy (ADT): (i) knowledge, self-efficacy (SE), and health beliefs about osteoporosis (OP); (ii) current engagement in healthy bone behaviours (HBBs). To explore the relationships between knowledge, SE, and health beliefs, and engagement in HBBs., Patients and Methods: 175 patients receiving ADT by injection completed questionnaires assessing current HBBs, OP knowledge, SE, and health beliefs (motivation, perceived susceptibility, and seriousness). Descriptive statistics and independent samples t-tests were used to assess relationships between knowledge, SE, health beliefs, and engagement in HBBs., Results: Only 38% of patients had undergone a dual X-ray absorptiometry scan in the past 2 years. OP knowledge was low (mean [sd, range] 9.6 [4.4, 0-19]) and perceived SE moderate (84.7 [24.5, 0-120]). Health motivation was fairly high (23.6 [3.1, 6-30]), but perceived susceptibility (16.8 [4.3]) and seriousness (16.8 [4.2]) of OP were low. Few patients met the recommendations for vitamin D intake (42%) and exercise (31%), and 15% were at risk of over-supplementation of calcium. Patients taking calcium supplements (P = 0.04), and meeting guidelines for vitamin D (P = 0.008) and for exercise (P = 0.002) had significantly greater knowledge than those who did not. Patients who were engaging in less than four of five HBBs had lower knowledge (P < 0.001) and health motivation (P = 0.01) than those who were engaging in four or all five HBBs., Conclusions: Most patients who are receiving ADT are not receiving appropriate screening, lack basic information about bone health, and are not engaging in the appropriate HBBs. These findings support the application of the Health Belief Model in this population: interventions that teach patients about the implications of bone loss, encourage proper uptake of HBBs, and promote feelings of SE could increase engagement in HBBs to prevent and manage bone loss., (© 2013 BJU International.)

Background: Patients with head and neck cancer experience complex posttreatment challenges. In a preliminary uncontrolled study, we evaluated the extent to which they can benefit from an interdisciplinary rehabilitation program., Methods: Twenty-seven patients completed an 8-week nutrition-rehabilitation program. Six-minute walk distance (6MWD), body weight, symptom burden, distress, and quality of life were assessed at baseline and at the end of the program., Results: Patients improved their mean 6MWD by 59 m (effect size = 0.8), and 78% of patients either maintained or increased their body weight. They also reported a clinically meaningful reduction in severity of insomnia, pain, weakness, anorexia, shortness of breath, depression and distress, and improvement in quality of life (effect sizes = 0.6-0.9). However, they reported no change in symptom interference with function., Conclusion: An interdisciplinary rehabilitation program may be beneficial to patients with head and neck cancer after treatment, but its effects should be evaluated in a controlled trial., (Copyright © 2012 Wiley Periodicals, Inc.)

Background and Aims: The diagnostic accuracy of capsule endoscopy has been suggested to be influenced by the direction of the passage in the intestine. It is currently unknown if a head-first or a tail-first orientation are equally common during the descent through the small bowel. The aim of the study was to identify the orientation of the capsule along the migration through the small bowel., Methods: Thirty capsule endoscopies were reviewed by an experienced observer. The direction of the passage through the pylorus and the ileoceccal valve was recorded for all the examinations. In addition, detailed review of the passage of the capsule in different segments of the small bowel was undertaken for all the capsules., Results: The capsule was significantly more likely to pass the pylorus head-first compared to tail-first (25 and 5 out of 30, respectively, OR 5, 95% CI 65-94%, P < 0.001). In 28/30 studies, the capsule exited the ileoceccal valve head-first (OR-14, 95% CI 77-99%, P < 0.001). In an immersion experiment, uneven distribution of weight of the capsule body was demonstrated with the head part (camera tip) being lighter than the tail part., Conclusions: The capsule endoscope usually passes through the pylorus and subsequent segments of the small bowel head-first. This observation suggests that the intestinal peristaltic physiology drives symmetrical bodies with their light part first. The principle of intestinal orientation by weight distribution may bear implications for capsules' design in the future.

Background & Aims: There are few data on risk of travel for patients with inflammatory bowel disease (IBD). We assessed rates of illness while traveling among patients with IBD., Methods: We performed a retrospective, case-controlled study of illnesses among 222 patients with IBD and 224 healthy individuals (controls) during 1099 total trips. Data were retrieved by structured questionnaires, personal interviews, and chart review., Results: Participants had 142 episodes of illness during the trips; 92% were enteric disease. An episode of illness occurred during 79/523 (15.1%) trips made by patients with IBD compared with 63/576 (10.9%) trips made by controls (odds ratio [OR], 1.44; 95% confidence interval [CI], 1.01-2.0; P = .04). However, this difference was mostly attributable to the increased incidence of illness among IBD patients traveling in industrialized countries. In contrast, the rate of illness among travelers to developing countries was similar among patients with IBD and controls (34/200, 17% vs 52/243, 21% of trips, respectively; P = .24). Moreover, numerically more controls that traveled to the tropics developed illness than travelers with IBD (43/135 vs 23/97, respectively; P = .18). In multivariate analysis, factors that increased risk for travel illness included frequent flares of IBD (OR, 1.9; 95% CI, 1.1-3.4; P = .02) and prior IBD-related hospitalizations (OR, 3.5; 95% CI, 1.3-9.3; P = .01); remission within 3 months before traveling reduced the risk for illness (OR, 0.3; 95% CI, 0.16-0.5; P < .001). Use of immunomodulatory drugs was not independently associated with risk of illness during travel., Conclusions: Patients with IBD have a higher rate of illness compared with controls during trips to industrialized countries, but not to developing or tropical regions. These findings indicate that most travel-associated illnesses stem from sporadic IBD flares rather than increased susceptibility to enteric infections., (Copyright © 2012 AGA Institute. Published by Elsevier Inc. All rights reserved.)

Background: A strategy to increase the number of size- and weight-appropriate organs and decrease the paediatric waiting list mortality is wider application of sectional orthotopic liver transplantation (OLT). These technical variants consist of living donor, deceased donor reduced and split allografts. However, these grafts have an increased risk of biliary complications. An unusual and complex biliary complication which can lead to graft loss is inadvertent exclusion of a major segmental bile duct. We present four cases and describe an algorithm to correct these complications., Methods: A retrospective review of the paediatric orthotopic liver transplantation database (2000-2010) at Washington University in St. Louis/St. Louis Children's Hospital was conducted., Results: Sixty-eight patients (55%) received technical variant allografts. Four complications of excluded segmental bile ducts were identified. Percutaneous cholangiography provided diagnostic confirmation and stabilization with external biliary drainage. All patients required interval surgical revision of their hepaticojejunostomy for definitive drainage. Indwelling biliary stents aided intra-operative localization of the excluded ducts. All allografts were salvaged., Discussion: Aggressive diagnosis, percutaneous decompression and interval revision hepaticojejunostomy are the main tenets of management of an excluded bile duct. Careful revision hepaticojejunostomy over a percutaneous biliary stent can result in restoration of biliary continuity and allograft survival., (© 2011 International Hepato-Pancreato-Biliary Association.)

Background: A family history of inflammatory bowel disease (IBD) is present in some ulcerative colitis (UC) patients. We aimed to investigate the familial occurrence of UC and its impact on disease severity., Methods: A structured questionnaire was distributed to patients with UC. Parameters pertaining to disease severity were compared for patients with or without positive family history of IBD., Results: The study group consisted of 168 UC patients with a total of 952 first degree relatives. Positive family history for IBD in a first degree relative was reported in 24 patients (14%). Six of the 336 parents (1.8%) had IBD (all with UC). There were 13 siblings with IBD (4 CD, 9 UC) out of 249 (5.4%). Seven of 376 (1.9%) offsprings had IBD (4 CD, 3 UC). Familial patients were more commonly females and have reported significantly more disease exacerbations than the sporadic group (17.7±15 versus 6.8±11, respectively, p=0.006). On multivariate analysis, familial disease was significantly and independently associated with both female sex (OR 4.1, 95% CI 1.1-14.9, p=0.04) and more exacerbations per year (annual OR 1.05, 95% CI 1.01-1.1, p=0.02). However, similar proportions of sporadic and familial patients wherever hospitalized, underwent colectomy or were treated by immune-suppressors., Conclusions: Familial occurrence of UC is not uncommon among Jewish patients in Israel. The familial-genetic component may preferentially influence disease occurrence among females, and is possibly associated with more disease flares although other parameters of disease severity do not seem to be impacted.

Background: Intestinal obstruction is a rare but serious complication after liver transplantation. Adhesions are the most common cause of obstruction in the nontransplantation setting; however, after pediatric liver transplantation unusual causes must be considered., Study Design: A prospectively maintained institutional database was analyzed for all reoperations for intestinal obstruction on pediatric liver allograft recipients from 1990 to 2009., Results: During the study period, 181 pediatric patients underwent liver transplantation at the study center. The most common indication for transplantation was biliary atresia. Seven patients required reoperation for intestinal obstruction. All 7 patients had abdominal operations before transplantation and 5 of 7 received reduced-size grafts. No patients had adhesive small bowel obstruction. The cause was right-sided diaphragmatic hernia in 4 and post-transplantation lymphoproliferative disorder (PTLD) in 3. Diaphragmatic hernia was demonstrated by chest radiograph in 3 of 4 patients. The fourth was taken to surgery with a presumptive diagnosis of intestinal obstruction and a diaphragmatic hernia was found at exploration. In patients with PTLD causing obstruction, 2 presented with an obstructing mass and the third presented with intussusception. Mean time to reoperation was 29 months after liver transplantation. Patients with diaphragmatic hernia presented earlier post-transplantation than those with PTLD (4.2 ± 2.4 months versus 59.3 ± 54.6 months, respectively; p = 0.0003, Fisher's exact test). Six patients are alive at a median follow-up of 5.8 years. One patient succumbed to recurrent B-cell lymphoma., Conclusions: Intestinal obstruction after pediatric liver transplantation is commonly related to what would conventionally be considered unusual causes. A high index of suspicion must be maintained and early operative therapy considered as obstruction because causes such as diaphragmatic hernia and PTLD are unlikely to resolve with conservative measures., (Copyright © 2011 American College of Surgeons. Published by Elsevier Inc. All rights reserved.)

Spatial chromatin organization is emerging as an important mechanism to regulate the expression of genes. However, very little is known about genome architecture at high-resolution in vivo. Here, we mapped the three-dimensional organization of the human Hox clusters with chromosome conformation capture (3C) technology. We show that computational modeling of 3C data sets can identify candidate regulatory proteins of chromatin architecture and gene expression. Hox genes encode evolutionarily conserved master regulators of development which strict control has fascinated biologists for over 25 years. Proper transcriptional silencing is key to Hox function since premature expression can lead to developmental defects or human disease. We now show that the HoxA cluster is organized into multiple chromatin loops that are dependent on transcription activity. Long-range contacts were found in all four silent clusters but looping patterns were specific to each cluster. In contrast to the Drosophila homeotic bithorax complex (BX-C), we found that Polycomb proteins are only modestly required for human cluster looping and silencing. However, computational three-dimensional Hox cluster modeling identified the insulator-binding protein CTCF as a likely candidate mediating DNA loops in all clusters. Our data suggest that Hox cluster looping may represent an evolutionarily conserved structural mechanism of transcription regulation.

Biliary complications in pediatric LT are important causes of morbidity and graft loss. We examined our recent pediatric LT experience to determine the outcome of post-LT biliary complications and their relationship to graft type. All initially isolated LTs performed at our institution between January 1, 2000 and August 20, 2007 were reviewed. Recipient data, donor type, graft survival, and biliary complications data were examined. Of 66 LTs, 32 patients received whole organ grafts, and 34 received partial grafts; 11 split, seven reduced size, and 16 live donors. Seventy-seven percent of patients had biliary reconstruction using a RYH. Overall, 17 (26%) developed biliary complications, and 15 were diagnosed within six months post-LT. Live donor and split allografts had more biliary complications than reduced size or whole allografts (50% and 36% vs. 0% and 16%, respectively). Seventy-one percent responded to percutaneous or endoscopic treatment. Five failed initial non-operative management and required reoperation (one retransplantation). These data suggest that biliary strictures occur most frequently in live donor and split allografts and that non-operative therapy is highly successful. Partial grafts are essential in pediatric LT, and a high clinical suspicion for biliary complications combined with aggressive and early diagnosis and therapy rarely results in graft loss.

The 60-kDa heat shock protein (mHsp60) is a vital cellular complex that mediates the folding of many of the mitochondrial proteins. Its function is executed in cooperation with the co-chaperonin, mHsp10, and requires ATP. Recently, the discovery of a new mHsp60-associated neurodegenerative disorder, MitCHAP-60 disease, has been reported. The disease is caused by a point mutation at position 3 (D3G) of the mature mitochondrial Hsp60 protein, which renders it unable to complement the deletion of the homologous bacterial protein in Escherichia coli (Magen, D., Georgopoulos, C., Bross, P., Ang, D., Segev, Y., Goldsher, D., Nemirovski, A., Shahar, E., Ravid, S., Luder, A., Heno, B., Gershoni-Baruch, R., Skorecki, K., and Mandel, H. (2008) Am. J. Hum. Genet. 83, 30-42). The molecular basis of the MitCHAP-60 disease is still unknown. In this study, we present an in vitro structural and functional analysis of the purified wild-type human mHsp60 and the MitCHAP-60 mutant. We show that the D3G mutation leads to destabilization of the mHsp60 oligomer and causes its disassembly at low protein concentrations. We also show that the mutant protein has impaired protein folding and ATPase activities. An additional mutant that lacks the first three amino acids (N-del), including Asp-3, is similarly impaired in refolding activity. Surprisingly, however, this mutant exhibits profound stabilization of its oligomeric structure. These results suggest that the D3G mutation leads to entropic destabilization of the mHsp60 oligomer, which severely impairs its chaperone function, thereby causing the disease.

Background: Double balloon enteroscopy is a new technique that enables deep intubation of the endoscope into the small bowel lumen. Through a channel in the endoscope, invasive procedures such as biopsy, polypectomy and hemostasis can be performed, avoiding the need for surgery., Objectives: To prospectively analyze our results of the first 124 DBEs performed since February 2007., Methods: The study group comprised all patients who underwent DBE at the Sheba Medical Center between February 2007 and February 2009. Recorded were the patients' demographic data, comorbidities, indications for the examination, results of previous non-invasive small bowel imaging (computed tomography enterography, capsule endoscopy, etc), investigation time, and results of the procedure including findings, endoscopic interventions, complications and pathological report., Results: A total of 124 procedures were performed in 109 patients. Of the 124 examinations, 57 (46%) were normal and 67 (54%) showed pathology. The main pathologies detected on DBE were polyps (14%), vascular lesions (17.6%) and inflammation (12%). Endoscopic biopsies and therapeutic interventions were required in 58 examinations (46%). A new diagnosis was established in 15% of patients, diagnosis was confirmed in 29% and excluded or corrected in 12%. One complication was observed: a post-polypectomy syndrome that was treated conservatively., Conclusions: DBE is a safe procedure and has a high diagnostic and therapeutic yield. Most of the examinations were performed under conscious sedation, and only a minority of patients required deeper sedation.

Monitoring CSA levels at two h after dosing (C2) has been shown effective in providing adequate CSA-based immunosuppression in clinical trials in adult transplant recipients, but there is limited data regarding C2 monitoring in pediatric transplant recipients. Given the differences in CSA pharmacokinetics between children and adults, a cohort of stable pediatric transplant recipients was converted from monitoring CSA trough (C0) to C2 levels, to establish the clinical utility and safety of C2 monitoring. After an abbreviated AUC(0-5) to establish baseline exposure, subsequent CSA dosing was adjusted based on C2 levels. Additional evaluation included monitoring for rejection, changes in CSA dose, toxicity, serum chemistries, and infection. Twelve heart transplant recipients were enrolled, with mean age 4.8 yr (range: 0.6-14.0). All patients received microemulsified CSA (Neoral((R)); Novartis Pharmaceuticals, East Hanover, NJ, USA) twice daily. Baseline CSA dose was 5.39 +/- 2.05 mg/kg/day (mean +/- s.d.), with mean C0 = 267 +/- 112, C2 = 1065 +/- 565, and AUC(0-5) = 3817 +/- 1435. Only seven participants showed clear CSA peak levels at two h, with five exhibiting delayed peaks at three to five h post-dose. These seven participants completed 48 wk of study, with mean CSA dose decreasing to 4.55 +/- 3.61 mg/kg/day, maintaining mean C2 599 +/- 211 (vs. target C2 = 800). No significant change in serum creatinine was observed, although GFR increased from 76.9 to 107.6 mL/min/1.73 m(2) (p = 0.11). Five patients failed to achieve target C2 levels (>800) during the first four wk, despite comparable AUC values, and were maintained on trough monitoring (C0). Mean systolic and diastolic blood pressures fell slightly, three minor infections were noted during the study period, and one episode of acute rejection occurred, despite stable CSA dosing. Nearly 50% of stable pediatric transplant recipients failed to achieve adequate peak C2 CSA levels during conversion from C0 to C2 monitoring. Age-dependent differences in CSA absorption and/or clearance pharmacokinetics may explain these findings.

Background: Simultaneous liver-lung transplantation is an infrequent but technically feasible procedure in patients with end-stage lung disease and advanced liver disease. We characterize the outcomes of pediatric patients who underwent this procedure at our institution., Methods: We performed a retrospective, case-control study and reviewed the medical records of all patients referred to our transplant program from its inception. Seven patients were listed for simultaneous liver-lung transplant. The five patients who survived to transplant were matched to 13 controls who underwent isolated bilateral sequential lung transplant for underlying diagnosis, age at time of transplant, gender, and era of transplant. Outcome measures included patient and graft survival, occurrence of bronchiolitis obliterans (BO), and episodes of rejection., Results: Of the five study patients who underwent liver-lung transplant, one died of multiorgan failure 11 days after transplant compared with 9 of 13 controls who died. The median survival for the study patients was 89 months (range, 0-112 months) compared with the controls, who had a median survival of 34 months (range, 0-118 months). The remaining four patients had bronchiolitis obliterans syndrome scores of 0 compared with 5 of 13 control patients (P=0.02). The rate of acute rejection per 100 patient days was 0.012 for the study patients compared with 0.11 for the controls (P=0.025)., Conclusions: Simultaneous liver-lung transplantation is a technically feasible procedure with excellent long-term outcomes. The surviving study subjects remain free from bronchiolitis obliterans syndrome. These results suggest that the transplanted liver may bestow immunologic privilege to the lung allograft.

Background and Goals: Video capsule endoscopy (VCE) enables visualization of the entire small bowel and can identify lesions that may go undetected by conventional endoscopy and radiography. In this study, we assessed whether patient's selection based on symptoms may increase the yield of VCE in the diagnosis of Crohn's disease (CD)., Study: Findings of 125 consecutive patients referred for VCE in whom CD may be suspected, were analyzed. Indications for VCE included iron-deficiency anemia, abdominal pain, diarrhea, or a combination of symptoms. Capsule endoscopy (CE) results were defined positive if 4 or more obvious clear ulcers, erosions, or a region with clear exudate and mucosal hyperemia and edema were identified., Results: One hundred twelve patients were included in the final analysis. Mean age of patients was 44+/-22 years and median follow-up 36+/-15 months. Findings on CE were considered compatible with a diagnosis of CD in 7 patients (6%). In general, CE yielded a diagnosis of CD in a very small portion of the patients (0% to 4%), except in patients undergoing the test for a combination of abdominal pain and diarrhea. In this group, findings suggestive of inflammatory bowel disease were encountered in one-third of the patients (P=0.002)., Conclusions: The greatest yield of CE in diagnosing CD is achieved in young patients who present with symptoms of abdominal pain plus diarrhea.

Background: Endoscopic ultrasound (EUS) has been shown to be a reliable tool for staging rectal cancer. Nevertheless, the accuracy of EUS after chemoradiation remains unclear; therefore the purpose of the present paper was to compare the accuracy of EUS staging for rectal cancer before and following chemoradiation., Methods: Patients with rectal cancer undergoing EUS staging were stratified into two groups. Group I consisted of 66 patients who underwent surgery following EUS staging without preoperative chemoradiation. Group II consisted of 25 patients who had EUS evaluation following chemoradiation. The EUS staging was compared to surgical/pathological staging., Results: The accuracy of the T staging for group I was 86% (57/66). Inaccurate staging was mainly associated with overstaging EUS T2 tumors. The accuracy of the N staging for group I was 71% (47/66). The accuracy of EUS for a composite T and N staging relevant to treatment decisions in group I was 91%. In group II, the accuracy of T and N staging was 72% (18/25) and 80% (20/25), respectively. Overstaging EUS T3 tumors accounted for most inaccurate staging. The EUS staging predicted post-chemoradiation T0N0 stage correctly in only 50% of cases., Conclusions: Preoperative staging of rectal cancer by EUS is a useful modality in determining the need for preoperative chemoradiation. The EUS T staging following chemoradiation appears to be less accurate. Detection of complete response may be insufficient for selecting patients for limited surgical intervention.

Background: Patients with iron deficiency anemia are subjected to multiple endoscopic and radiologic examinations of the GI tract. If negative, some of the examinations are repeated, occasionally with positive findings. The diagnostic yield of a second capsule endoscopy in such patients is unknown. The aim of the current study was to assess the diagnostic yield of a second capsule endoscopy in patients with significant iron deficiency anemia and a previous negative evaluation., Methods: Twenty patients with iron deficiency anemia (Hb < 10 g/dL) were enrolled. All had at least one normal evaluation of the GI tract, including capsule endoscopy. A second capsule endoscopy examination was offered to all patients., Results: The time between the first and the second capsule endoscopy ranged from 2 months to 1 year. Depending on the nature of an abnormality and its relevance to blood loss, the findings were classified as positive, suspicious, clinically irrelevant, or negative. In 7 patients, the second capsule endoscopy disclosed findings that were classified as either positive or suspicious findings, including arteriovenous malformations (2), flat polypoid lesion (1), edematous inflamed mucosa (1), erosions (1), and hemorrhagic gastritis (1). Lesions were located in the small intestine (5), the stomach (1), and the cecum (1). Based on the findings of the second capsule endoscopy, therapy was changed in two patients (10%)., Conclusions: A second capsule endoscopy should be considered for patients with severe iron deficiency anemia and negative initial evaluation.

The molecular mechanisms that regulate basal or background entry of divalent cations into mammalian cells are poorly understood. Here we describe the cloning and functional characterization of a Ca2+- and Mg2+-permeable divalent cation channel, LTRPC7 (nomenclature compatible with that proposed in ref. 1), a new member of the LTRPC family of putative ion channels. Targeted deletion of LTRPC7 in DT-40 B cells was lethal, indicating that LTRPC7 has a fundamental and nonredundant role in cellular physiology. Electrophysiological analysis of HEK-293 cells overexpressing recombinant LTRPC7 showed large currents regulated by millimolar levels of intracellular Mg.ATP and Mg.GTP with the permeation properties of a voltage-independent divalent cation influx pathway. Analysis of several cultured cell types demonstrated small magnesium-nucleotide-regulated metal ion currents (MagNuM) with regulation and permeation properties essentially identical to the large currents observed in cells expressing recombinant LTRPC7. Our data indicate that LTRPC7, by virtue of its sensitivity to physiological Mg.ATP levels, may be involved in a fundamental process that adjusts plasma membrane divalent cation fluxes according to the metabolic state of the cell.

Cutaneomuscular reflex (CMR) responses and motor unit synchronisation have been recorded to investigate possible reorganisation of central nervous pathways in six healthy adults learning a novel skill with the non-dominant hand. Multi-unit surface EMG signals were recorded from first dorsal interosseous (1DI) and abductor digiti minimi (ADM) muscles during sustained flexion of the index finger and abduction of the little finger (flex/ab) and during bilateral controlled finger abduction (ab/ab). CMR responses were elicited by concomitant stimulation of the digital nerves of the index finger at 2.5 times threshold for perception. For training purposes the subject practised the novel flex/ab task phasically at a rate of 1 Hz for 10 min daily for 12 days. Cortically mediated components of the CMR responses recorded from 1DI and ADM were significantly larger in both muscles during training. Taking the data for all subjects together, when subjects performed sustained flex/ab (novel task), the size of the I1 and E2 components were increased by 25 and 55% in IDI and 192 and 167% in ADM (Mann-Whitney U-test, P<0.05). Corresponding values for sustained ab/ab were more modest: 31 and 16% in 1DI and 88 and 54% in ADM. Changes in the size of the spinal E1 component were not the same in each muscle: values for sustained flex/ab and ab/ab were 33 and 31% smaller in 1DI and 89 and 59% larger in ADM. No significant changes were found in the amount of synchrony of motor unit firing between 1DI and ADM when subjects performed either task during training. These results suggest that learning a new motor skill produces changes which take place predominantly in the cortical pathways of the CMR and these may be due to changed connectivity within motor and/or sensory cortex which has previously been shown in the monkey.

Objective: To evaluate the virological efficacy and safety of quadruple therapy with two nucleoside analogues and ritonavir (400 mg twice daily) plus indinavir (400 mg twice daily) combination in antiretroviral therapy-naive patients., Design and Methods: An open-label, uncontrolled multicentre trial. Antiretroviral therapy-naive patients (n = 90) with high median baseline HIV RNA levels of 220,000 copies/ml (range, 36,000-2,943,000 copies/ml) and median CD4 cell count of 189 x 10(6)/l (range, 4-656 x 10(6)/l) were started on a twice daily regimen of either zidovudine/lamivudine (49%), stavudine/lamivudine (38%) or stavudine/didanosine (13%) plus ritonavir 400 mg twice daily and indinavir 400 mg twice daily combination therapy. CD4 cell counts and HIV RNA were determined at weeks 0, 4, 8, 12, 16, 20, and 24. Statistical analysis was performed on treatment as well as intent-to-treat, where missing values were accounted for as failure., Results: In the intent-to-treat analysis at week 24, the proportion of patients with HIV RNA of < 500 copies/ml, and < 80 copies/ml was 86.7% and 71.1%, respectively. In the on-treatment analysis at week 24, 80.0% of patients had undetectable viral load in the ultrasensitive assay (< 80 copies/ml; n = 80). The quadruple therapy was well tolerated except for mild diarrhoea, initial nausea and increased triglyceride levels. Treatment was stopped in seven (7.7%) patients because of adverse events and three (3.3%) were lost to follow-up., Conclusions: Our preliminary data suggest that the protease inhibitor combination ritonavir/indinavir plus double nucleoside therapy appears to be effective and safe in short-term treatment (up to 24 weeks).

Astrocytic proliferation and hypertrophy (astrogliosis) are associated with neuronal injury. However, neither the temporal nor the spatial relationship between astrocytes and injured neurons is clear, especially in neurodegenerative diseases. We investigated these questions in a mouse amyotrophic lateral sclerosis (ALS) model. The initial increase in astrogliosis coincided with the onset of clinical disease and massive mitochondrial vacuolation in motor neurons. After disease onset, astrogliosis increased further in parallel with the number of degenerating motor neurons. Examination of individual astrocytes by three-dimensional reconstruction revealed that astrocytes extended their processes toward, wrapped around, and sometimes penetrated vacuoles derived from neuronal mitochondria. These results show a close temporal correlation between the onset of neuronal degeneration and the beginning of astrogliosis in this neurodegenerative disease and reveal a novel spatial relationship that is consistent with the view that astrocytes play an active role in the neuronal degeneration process., (Copyright 1999 Wiley-Liss, Inc.)

Background: Signals from the B-cell antigen receptor (BCR) help to determine B-cell fate, directing either proliferation, differentiation, or growth arrest/apoptosis. The protein tyrosine phosphatase SHP-1 is known to regulate the strength of BCR signaling. Although the B-cell co-receptor CD22 binds SHP-1, B cells in CD22-deficient mice are much less severely affected than those in SHP-1-deficient mice, suggesting that SHP-1 may also regulate B-cell signaling by affecting other signaling molecules. Moreover, direct substrates of SHP-1 have not been identified in any B-cell signaling pathway., Results: We identified the B-cell transmembrane protein CD72 as a new SHP-1 binding protein and as an in vivo substrate of SHP-1 in B cells. We also defined the binding sites for SHP-1 and the adaptor protein Grb2 on CD72. Tyrosine phosphorylation of CD72 correlated strongly with BCR-induced growth arrest/apoptosis in B-cell lines and in primary B cells. Preligation of CD72 attenuated BCR-induced growth arrest/death signals in immature and mature B cells or B-cell lines, whereas preligation of CD22 enhanced BCR-induced growth arrest/apoptosis., Conclusions: We have identified CD72 as the first clear in vivo substrate of SHP-1 in B cells. Our results suggest that tyrosine-phosphorylated CD72 may transmit signals for BCR-induced apoptosis. By dephosphorylation CD72. SHP-1 may have a positive role in B-cell signaling. These results have potentially important implications for the involvement of CD72 and SHP-1 in B-cell development and autoimmunity.

The protein tyrosine phosphatase SHP-1 is a critical regulator of macrophage biology, but its detailed mechanism of action remains largely undefined. SHP-1 associates with a 130-kDa tyrosyl-phosphorylated species (P130) in macrophages, suggesting that P130 might be an SHP-1 regulator and/or substrate. Here we show that P130 consists of two transmembrane glycoproteins, which we identify as PIR-B/p91A and the signal-regulatory protein (SIRP) family member BIT. These proteins also form separate complexes with SHP-2. BIT, but not PIR-B, is in a complex with the colony-stimulating factor 1 receptor (CSF-1R), suggesting that BIT may direct SHP-1 to the CSF-1R. BIT and PIR-B bind preferentially to substrate-trapping mutants of SHP-1 and are hyperphosphorylated in macrophages from motheaten viable mice, which express catalytically impaired forms of SHP-1, indicating that these proteins are SHP-1 substrates. However, BIT and PIR-B are hypophosphorylated in motheaten macrophages, which completely lack SHP-1 expression. These data suggest a model in which SHP-1 dephosphorylates specific sites on BIT and PIR-B while protecting other sites from dephosphorylation via its SH2 domains. Finally, BIT and PIR-B associate with two tyrosyl phosphoproteins and a tyrosine kinase activity. Tyrosyl phosphorylation of these proteins and the level of the associated kinase activity are increased in the absence of SHP-1. Our data suggest that BIT and PIR-B recruit multiple signaling molecules to receptor complexes, where they are regulated by SHP-1 and/or SHP-2.

When incubated with [3H]myristate or [3H]palmitate, LSTRA cells, a murine T cell line, incorporated radiolabel into a protein of 95 kDa as analyzed by SDS-polyacrylamide gel electrophoresis. This dually acylated protein was identified as the transferrin receptor by immunoprecipitation with a monoclonal anti-transferrin receptor antibody. Acylation of the transferrin receptor was posttranslational and occurred via ester or thioester linkages. Analysis of radiolabeled transferrin receptor protein from [3H]myristate-labeled cells by acid hydrolysis followed by thin layer chromatography revealed the exclusive presence of [3H]myristate. Labeled transferrin receptor protein from [3H]palmitate-labeled cells contained predominantly [3H]stearate and smaller amounts of [3H]palmitate. This is in contrast to the protein-tyrosine kinase p56lck, which in [3H]palmitate-treated LSTRA cells, incorporated primarily [3H]palmitate. An analog of myristic acid, 5-nonanyloxyfuran-2-carboxylic acid, inhibited the incorporation of [3H]myristate, but not [3H]palmitate or [3H]stearate into transferrin receptor protein, suggesting that these acylation events are distinct. These studies indicate that the murine transferrin receptor is acylated posttranslationally with myristate, palmitate and stearate and suggest that more than one acyltransferase activity is responsible for its acylation.

N-Myristoylation of p56lck, a member of the Src family of protein-tyrosine kinases, is essential for its proper targeting to the plasma membrane. 2-Hydroxymyristic acid (HMA) is an analog of myristic acid that becomes metabolically activated in cells to form 2-hydroxymyristoyl-CoA, a potent inhibitor of myristoyl-CoA:protein N-myristoyltransferase (NMT), the enzyme that catalyzes protein N-myristoylation [Paige, L. A., Zheng, G.-q., DeFrees, S. A., Cassady, J. M., & Geahlen, R. L. (1990) Biochemistry 29, 10566]. In the presence of HMA, LSTRA cells, which overexpress p56lck, synthesized nonmyristoylated p56lck, which displayed a reduced electrophoretic mobility on SDS-polyacrylamide gels identical to that of a nonmyristoylated Gly2-->Ala2 mutant of p56lck. Treatment with myristic acid, 2-hydroxypalmitic acid, or 2-fluoromyristic acid did not result in the synthesis of nonmyristoylated p56lck. In contrast to the membrane-associated, myristoylated p56lck, nonmyristoylated p56lck was cytosolic. Although nonmyristoylated p56lck retained tyrosine kinase activity, it was not labeled in vivo with [32P]orthophosphate, indicating that a change in subcellular location altered its state of phosphorylation. A pulse-chase analysis revealed that cytosolic, nonmyristoylated p56lck was less stable than the myristoylated enzyme. In cell lines that do not overexpress p56lck, HMA treatment resulted in a reduction in the levels of both newly synthesized and total p56lck. Treatment of CD4+ cells with HMA caused a corresponding decrease in the amount of CD4-associated p56lck. Thus, chemical inhibition of protein N-myristoylation with HMA is an effective method for reducing the amount of p56lck available at the plasma membrane for signal transduction.

The myristoylated protein-tyrosine kinase, p56lck, is expressed predominantly in T cells where it is believed to play a role in T cell activation. We observed a 56-kDa protein that became metabolically labeled in intact T lymphoid cells that were incubated with either [3H]myristate or [3H]palmitate. This protein was identified as p56lck based on its specific immunoprecipitation with polyclonal antisera to p56lck, by induction of a shift in its electrophoretic mobility following treatment of cells with 12-O-tetradecanoylphorbol-13-acetate and by co-chromatography with p56lck on protamine-agarose. Characterization of the two acylation events revealed that, in contrast to the p56lck-associated radioactivity from [3H]myristate-labeled cells, the p56lck-associated radioactivity from [3H]palmitate-labeled cells was susceptible to cleavage by neutral hydroxylamine and was not blocked by inhibitors of protein synthesis. Pulse-chase analyses revealed that the labeling of p56lck with [3H]palmitate, but not [3H]myristate, was reversible. The presence of covalently attached palmitate on p56lck from [3H]palmitate-labeled cells was verified by thin-layer chromatography following acid hydrolysis of the acylated protein. 2-Hydroxymyristate, which is metabolically activated to form a potent inhibitor of protein myristoylation, specifically inhibited the acylation of p56lck with [3H]myristate without affecting its labeling with [3H]palmitate. These studies indicate that p56lck is both a cotranslationally myristoylated and post-translationally palmitoylated protein.

This study was designed to determine which of three quality assessment methods most validly identifies deficient care. Process criteria were developed to assess outpatient care for urinary tract infection using each of three methods: a limited "list" of seven criteria, an extensive "list" of 40 criteria, and a criteria map (CM) which uses branching logic to identify applicable criteria according to the specific needs of each case. Defining deficiency as compliance with less than 60 percent of criteria, the extensive list found all 66 cases deficient; the limited list, 27 (41.0 percent); and the CM system, 15 (22.7 percent). After excluding the extensive list because of its nondiscrimination, 23 discrepancies in rating remained between the limited list and the CM. Ten physicians unaware of the results reviewed all 23 cases. In 12 of these 23 cases, at least seven of the ten physicians preferred the rating of one method over another; the CM assessment was preferred in 11 of the 12 cases (P less than .01). Criteria maps, providing a patient-specific approach, offer a more valid assessment of medical care than either the extensive or limited list.

Eighty-four cataractous eyes of 52 consecutive patients and 81 normal eyes of 48 subjects were evaluated with a commercially availability glare tester. Tests results were compared to visual acuity determinations in the refracting lane for ability to predict vision outdoors under daylight conditions. Among cataract patients, predictability of the two methods of measurement was not significantly different when subjects faced away from the sun. Glare scores were significantly better predictors (nearly twice as predictive) of outdoor visual acuity among cataract subjects facing the sun. These results demonstrate that measurement of sensitivity to glare is an important clinical procedure for evaluating functional visual disability in cataract patients under glare-producing conditions.

A simple, inexpensive, readily available, easy-to-service clinical glare tester combines an audioviewer able-top projector with a series of specially made slides. Each slide supplies a constant glare source and a variable-contrast, central target. The device was evaluated on 161 normal subjects, 144 patients with cataracts, and 110 aphakic patients whose visual acuity was corrected with spectacles or intraocular lenses. Results showed that aphakic patients, irrespective of the mode of correction, were statistically more glare sensitive than normal phakic patients. Further results confirmed earlier studies that demonstrated that glare sensitivity also increased with age in normal eyes. Broad potential exists for using this method of glare testing in industry, transportation media, and medicolegal and and clinical ophthalmologic studies.

A private clinic was established in Los Angeles in late 1972 to perform acupuncture. Most individuals seeking treatment were middle-aged and well educated. All had considerable experience with traditional medical care for their problem. The principal reason for seeking care was relief from chronic pain. Thirteen per cent had received other nontraditional forms of therapy; over one-third indicated that they sought acupuncture as a "last resort." The results of certain psychological tests taken by these patients were within the range of normal. On completion of six or more treatments, 74 per cent said they had been helped; 54 per cent of those followed for three months and 24 per cent contacted one year later were improved. A subgroup of patients who received only temporary improvement had significantly different scores on psychological scales suggesting the results noted in this group may have been due to a "placebo" effect. A significant proportion of those who dropped out of treatment did so because they had achieved the desired benefits.

Survey responses showed little change in dental office staffing and paperwork as a result of the UAW/auto industry program. Only slight differences between respondents in the four states were apparent; except for Michigan, where the greatest increase in staff and additional clerical hours was reported. The differences that did exist might be related to the volume of UAW/auto industry business in each area and the dentists' opinions of the carriers they relate to most frequently. The responses did not reveal a clear indication of success or failure of the plan by virture of any category evaluated.

Diallyl thiosulfinate (allicin) is the agent found in garlic which is responsible for the antibacterial and antifungal activity of extracts of this plant. The effect of bacteriostatic concentrations of allicin (0.2 to 0.5 mM) on the growth of Salmonella typhimurium revealed a pattern of inhibition characterized by: (i) a lag of approximately 15 min between addition of allicin and onset of inhibition, (ii) a transitory inhibition phase whose duration was proportional to allicin concentration and inversely proportional to culture density, (iii) a resumed growth phase which showed a lower rate of growth than in uninhibited controls, and (iv) an entry into stationary phase at a lower culture density. Whereas DNA and protein syntheses showed a delayed and partial inhibition by allicin, inhibition of RNA synthesis was immediate and total, suggesting that this is the primary target of allicin action.