Translocation of a gut pathobiont drives autoimmunity in mice and humans.

Despite multiple associations between the microbiota and immune diseases, their role in autoimmunity is poorly understood. We found that translocation of a gut pathobiont, Enterococcus gallinarum, to the liver and other systemic tissues triggers autoimmune responses in a genetic background predisposing to autoimmunity. Antibiotic treatment prevented mortality in this model, suppressed growth of E. gallinarum in tissues, and eliminated pathogenic autoantibodies and T cells. Hepatocyte-E. gallinarum cocultures induced autoimmune-promoting factors. Pathobiont translocation in monocolonized and autoimmune-prone mice induced autoantibodies and caused mortality, which could be prevented by an intramuscular vaccine targeting the pathobiont. E. gallinarum-specific DNA was recovered from liver biopsies of autoimmune patients, and cocultures with human hepatocytes replicated the murine findings; hence, similar processes apparently occur in susceptible humans. These discoveries show that a gut pathobiont can translocate and promote autoimmunity in genetically predisposed hosts.

Calcium signalling links MYC to NUAK1.

NUAK1 is a member of the AMPK-related family of kinases. Recent evidence suggests that NUAK1 is an important regulator of cell adhesion and migration, cellular and organismal metabolism, and regulation of TAU stability. As such, NUAK1 may play key roles in multiple diseases ranging from neurodegeneration to diabetes and metastatic cancer. Previous work revealed a crucial role for NUAK1 in supporting viability of tumour cells specifically when MYC is overexpressed. This role is surprising, given that NUAK1 is activated by the tumour suppressor LKB1. Here we show that, in tumour cells lacking LKB1, NUAK1 activity is maintained by an alternative pathway involving calcium-dependent activation of PKCα. Calcium/PKCα-dependent activation of NUAK1 supports engagement of the AMPK-TORC1 metabolic checkpoint, thereby protecting tumour cells from MYC-driven cell death, and indeed, MYC selects for this pathway in part via transcriptional regulation of PKCα and ITPR. Our data point to a novel role for calcium in supporting tumour cell viability and clarify the synthetic lethal interaction between NUAK1 and MYC.

Identification of LDH-A as a therapeutic target for cancer cell killing via (i) p53/NAD(H)-dependent and (ii) p53-independent pathways.

Most cancer cells use aerobic glycolysis to fuel their growth. The enzyme lactate dehydrogenase-A (LDH-A) is key to cancer's glycolytic phenotype, catalysing the regeneration of nicotinamide adenine dinucleotide (NAD(+)) from reduced nicotinamide adenine dinucleotide (NADH) necessary to sustain glycolysis. As such, LDH-A is a promising target for anticancer therapy. Here we ask if the tumour suppressor p53, a major regulator of cellular metabolism, influences the response of cancer cells to LDH-A suppression. LDH-A knockdown by RNA interference (RNAi) induced cancer cell death in p53 wild-type, mutant and p53-null human cancer cell lines, indicating that endogenous LDH-A promotes cancer cell survival irrespective of cancer cell p53 status. Unexpectedly, however, we uncovered a novel role for p53 in the regulation of cancer cell NAD(+) and its reduced form NADH. Thus, LDH-A silencing by RNAi, or its inhibition using a small-molecule inhibitor, resulted in a p53-dependent increase in the cancer cell ratio of NADH:NAD(+). This effect was specific for p53(+/+) cancer cells and correlated with (i) reduced activity of NAD(+)-dependent deacetylase sirtuin 1 (SIRT1) and (ii) an increase in acetylated p53, a known target of SIRT1 deacetylation activity. In addition, activation of the redox-sensitive anticancer drug EO9 was enhanced selectively in p53(+/+) cancer cells, attributable to increased activity of NAD(P)H-dependent oxidoreductase NQO1 (NAD(P)H quinone oxidoreductase 1). Suppressing LDH-A increased EO9-induced DNA damage in p53(+/+) cancer cells, but importantly had no additive effect in non-cancer cells. Our results identify a unique strategy by which the NADH/NAD(+) cellular redox status can be modulated in a cancer-specific, p53-dependent manner and we show that this can impact upon the activity of important NAD(H)-dependent enzymes. To summarise, this work indicates two distinct mechanisms by which suppressing LDH-A could potentially be used to kill cancer cells selectively, (i) through induction of apoptosis, irrespective of cancer cell p53 status and (ii) as a part of a combinatorial approach with redox-sensitive anticancer drugs via a novel p53/NAD(H)-dependent mechanism.

Anterosuperior anchoring myringoplasty technique for anterior and subtotal perforations.

Anterior and subtotal tympanic perforations are difficult perforations to repair. We used the anterosuperior anchoring technique to repair 105 of those perforations. Our technique utilizes a large temporalis fascia graft using an underlay technique with due emphasis on anterosuperior anchorage. The success rates in paediatric myringoplasty for consultant solo (J.R.K.) and trainees under supervision were 100% and 85.7%, respectively. The overall success rates in adult myringoplasty for consultant solo and trainees under supervision were 93.8% and 82.4%, respectively. Analysis of the 82 successful cases showed statistically significant improvement in air conduction thresholds after myringoplasty at 500 Hz (P < 0.01), 1 kHz (P < 0.05), 2 kHz (P < 0.01) and 4 kHz (P < 0.01). Our study showed that the anterosuperior anchoring technique produced excellent results in the repair of challenging anterior and subtotal perforations in both adults and children.

Major ear surgery in a paediatric day care unit.

Abstract Traditionally major ear surgery in children has been regarded as an in-patient procedure. Evidence from the USA for adults, however, concludes that it is both safe and effective to perform many major ear procedures as day cases. We have been carrying out major ear operations on children as day cases routinely for six years in a dedicated children's day unit and examined our data to find out whether it was both safe and feasible to perform major ear surgery in children on a day-case basis. As our main outcome measure we used the rate of unplanned admissions. We found that the unplanned admission rate for surgery, excluding mastoid surgery, was 6.7 per cent and that procedures such as myringoplasty, ossiculoplasty, bilateral pinnaplasty, meatoplasty and tympanotomy with excision of cholesteatoma, were eminently suitable for day surgery.

Tobacco use by Massachusetts public college students: long term effect of the Massachusetts Tobacco Control Program.

OBJECTIVE: To assess tobacco use among Massachusetts public college students and compare students who attended high school in Massachusetts and were exposed to the Massachusetts Tobacco Control Program (MTCP) with students who attended high school outside Massachusetts and were unexposed to the programme. DESIGN: Analysis of the 1999 Massachusetts College Alcohol Survey. SETTING: Four year public colleges and universities in Massachusetts (n = 11). SUBJECTS: 1252 randomly selected students (response rate 56%). MAIN OUTCOME MEASURES: Self report of current (past 30 day), past year, and lifetime use of cigarettes, cigars, and smokeless tobacco. RESULTS: One third of students had used a tobacco product in the past month and 46.4% had used tobacco in the past year. Cigarettes accounted for most of this tobacco use. Total tobacco use was higher among males than females but cigarette smoking did not differ by sex. Tobacco use was lower among athletes and higher among students who used alcohol or marijuana. Current tobacco use was lower among public college students who had attended high school in Massachusetts compared with those who attended high school in another state (31.5% v 42.6%, p = 0.006). This difference persisted after adjustment for age, sex, race, parental education, and students' college residence (adjusted odds ratio (OR) 0.67, 95% confidence interval (CI) 0.46 to 0.97, p = 0.034). CONCLUSION: Tobacco use is common among Massachusetts public college students. Students who were exposed to the MTCP during high school are less likely to use tobacco than their peers who were not exposed to this programme. The MTCP may have reduced tobacco use among this group of young adults.

Reliability of the Problem Oriented Screening Instrument for Teenagers (POSIT) in adolescent medical practice.

PURPOSE: To determine the internal consistency and 1-week test-retest reliability of the Problem Oriented Screening Instrument for Teenagers (POSIT) among adolescent medical patients. METHODS: A research assistant administered the POSIT to a consecutive sample of 15- to 18-year-old patients arriving for routine medical care at a hospital-based adolescent medical practice. Each subject returned for a retest 1 week later. Internal consistency for each scale on test and retest was calculated using Cronbach alpha, and 1-week test-retest reliability by the intraclass correlation coefficient (r) and the kappa coefficient (kappa). RESULTS: The Substance Use/Abuse, Mental Health Status, Educational Status, and Aggressive Behavior/Delinquency scales had favorable alpha scores (>.70). Others, including Physical Health Status, had lower alpha scores. High intraclass correlation coefficients were found for all 10 POSIT scales (r =.72 to.88), although (r) was lower for males on two of the scales. Kappa coefficients for all scales indicated good reproducibility beyond chance (kappa =.42 to.73). CONCLUSIONS: This study provided supportive evidence for the reliability of the POSIT in primary care medical settings, although some POSIT scales could likely be improved. The 20- to 30-min administration time is most practical in settings that are dedicated to adolescent medicine, and computerized administration and scoring are needed.

The role of the primary care provider in preventing and treating alcohol problems in adolescents.

Adolescents use alcohol more frequently and heavily than all other illicit drugs combined. Given the myriad health, developmental, and social problems associated with alcohol use, it is not surprising that the American Medical Association's Guidelines for Adolescent Preventive Services recommends that adolescents be asked annually about their use of alcohol, and those who report any use during the past year should be assessed further. However, routine alcohol screening of adolescents in primary care and emergency medical settings is not universally applied. In March 2000, the National Institute on Alcohol Abuse and Alcoholism of the National Institutes of Health and the Agency for Health care Research and Quality jointly sponsored a meeting entitled The Expanding Role of Primary Care in the Prevention and Treatment of Alcohol-Use Disorders. The purpose of the meeting was to bring together researchers, policymakers, clinicians, insurance providers, and medical education specialists to determine the best approaches to increase the involvement of primary care physicians and other health care professionals in screening and intervening for alcohol problems in their patients. The National Institute on Alcohol Abuse and Alcoholism and Agency for Health care Research and Quality believe that the evidence of efficacy for primary care involvement is compelling and are working together to promote the translation of these findings into clinical practice. The following article summarizes what is currently known about adolescent alcohol use and how it can be addressed in primary care settings. It provided the background for the meeting's focus on adolescent issues.

Development of a Bright Futures curriculum for pediatric residents.

OBJECTIVE: To develop a standardized case-based curriculum for pediatric residents on child growth, development, behavior, and adolescent medicine that incorporates the Bright Futures health supervision guidelines. DESIGN: This project included a needs assessment, development of a list of important topics, writing and revising of standardized cases, formative evaluation of cases, and efficacy pilot testing of 2 cases. SETTING: A large pediatric teaching hospital continuity clinic. PARTICIPANTS: Pediatric residents, fellows, and faculty. INTERVENTIONS: Preparation of standardized cases, facilitator training, and resident-led teaching conferences. OUTCOME MEASURES: Learner and facilitator evaluation forms and two 10-item diagnostic skills assessments. RESULTS: During the project, faculty-fellow teams wrote 29 case-teaching modules. All participants gave high ratings to cases, and resident facilitators reported increased comfort with the case discussion method. Resident learners' ability to accurately interpret developmental screening tests and growth charts improved following sessions on those topics. CONCLUSIONS: Further evaluation is required, but these standardized cases appear promising for teaching pediatric residents. The curriculum is now freely available to faculty nationwide.

Reliabilities of short substance abuse screening tests among adolescent medical patients.

OBJECTIVE: To determine the internal consistency and 1-week test-retest reliability of the Simple Screening Instrument for Alcohol and Other Drug Abuse (SSI-AOD), the CAGE-AA (CAGE questions adapted for adolescents), and 4 modified items from the Drug and Alcohol Problem QuickScreen (DAP-4) among adolescents. METHODS: Fifteen- to 18-year-old medical patients (n = 173) completed screening tests during a routine medical visit and then again 1 week later. Internal consistency for each test and retest was calculated using Cronbach's alpha, and 1-week test-retest reliability was calculated by using Winer's unbiased estimate of the intraclass correlation coefficient (r). RESULTS: The SSI-AOD has good internal consistency (alpha =.83) and the CAGE-AA questions acceptable internal consistency (alpha =.60). Alpha varied with gender and race, and item analysis indicated the CAGE-AA test could be improved. As expected, the DAP-4 had a lower alpha score (.46). All screening instruments studied had high 1-week test-retest reliabilities (range r =.82-.90). CONCLUSIONS: The SSI-AOD is a reliable substance abuse screening instrument among adolescent medical patients. The CAGE-AA questions must be further revised and tested before their use can be recommended. The DAP-4 questions are likely measuring different, but important, constructs.

A comprehensive analysis of protein-protein interactions in Saccharomyces cerevisiae.

Two large-scale yeast two-hybrid screens were undertaken to identify protein-protein interactions between full-length open reading frames predicted from the Saccharomyces cerevisiae genome sequence. In one approach, we constructed a protein array of about 6,000 yeast transformants, with each transformant expressing one of the open reading frames as a fusion to an activation domain. This array was screened by a simple and automated procedure for 192 yeast proteins, with positive responses identified by their positions in the array. In a second approach, we pooled cells expressing one of about 6,000 activation domain fusions to generate a library. We used a high-throughput screening procedure to screen nearly all of the 6,000 predicted yeast proteins, expressed as Gal4 DNA-binding domain fusion proteins, against the library, and characterized positives by sequence analysis. These approaches resulted in the detection of 957 putative interactions involving 1,004 S. cerevisiae proteins. These data reveal interactions that place functionally unclassified proteins in a biological context, interactions between proteins involved in the same biological function, and interactions that link biological functions together into larger cellular processes. The results of these screens are shown here.

Prevalence of alcohol problems among pediatric residents.

OBJECTIVE: To measure the prevalence of alcohol-related problems among pediatric trainees. METHODS: An alcoholism screening test was administered anonymously to participants at a mandatory substance abuse education and prevention program. SETTING: A large urban pediatric residency training program. SUBJECTS: One hundred fifteen pediatric residents attended the program during 3 consecutive years (1996-1998). Eighty-five (74%) screening tests were returned and 81 (70%) were analyzed MAIN OUTCOME MEASURE: The 25-item Michigan Alcoholism Screening Test (MAST). Differential MAST cutpoints have been established to "suggest" or "indicate" a lifetime diagnosis of alcoholism. RESULTS: Twelve residents (15%) had scores suggestive and 6 (7%) indicative of alcoholism. Twenty-eight (35%) admitted to having alcohol-associated amnesia (blackouts), 13 (16%) to "feeling bad" about their drinking, 9 (11%) to drinking before noon, 6 (7%) to getting into fights when drunk, and 2 (2%) to alcohol-related marital problems. However, only 1 (1%) had gone to anyone for help and none admitted to alcohol-related problems at work. CONCLUSIONS: These screening data suggest that alcohol abuse and related problems exist among pediatric trainees at troubling rates. While more than one third of the trainees had experienced a serious consequence from heavy drinking, only 1 had gone for help and problems were not apparent at work. Greater emphasis should be placed on alcohol prevention and early intervention programs as a routine part of pediatric training.

A new brief screen for adolescent substance abuse.

OBJECTIVE: To develop a brief alcohol and other drug (AOD) screening test for adolescents. METHODS: A 9-item test was constructed by combining and modifying items from several AOD assessments, and administered concurrently with the Personal Involvement With Chemicals Scale (PICS), the criterion standard. SETTING: A hospital-based adolescent clinic. SUBJECTS: Fourteen- to 18-year-old patients consecutively arriving for routine medical care who were known to have used AOD. MEASURES: Internal consistency of the 9 items was calculated using the Cronbach alpha. The relationship between the brief screen and PICS raw score was determined by stepwise linear regression analysis. The PICS T score has been shown to correctly classify substance abuse treatment need as no treatment (T<35), brief office intervention (T = 35-40), outpatient or short-term treatment (T = 41-54), and inpatient or long-term treatment (T> or =55). Sensitivity and specificity rates for predicting a PICS T score of 55 or higher were calculated from 2 x 2 tables. RESULTS: Ninety-nine adolescents were tested (70.7% female, 36.4% black, 32.3% white, 19.2% Hispanic, mean age, 16.3 years). The 9 items had good internal consistency (Cronbach alpha = .79). Stepwise linear regression analysis identified 6 items whose total combined score was highly correlated with PICS (Pearson r = 0.84, P<.01). This model correctly classified 86% of subjects according to the PICS criteria. Two or more yes answers had a sensitivity of 92.3% and specificity of 82.1% for intensive AOD treatment need. The 6 items were arranged into a mnemonic (CRAFFT). CONCLUSIONS: Further research must confirm the test's psychometric properties in a general clinic population. However, CRAFFT seems promising as a brief AOD screening test.

Recent patterns of use and associated risks of illicit drug use in adolescents.

Addressing adolescent substance abuse presents a tremendous challenge to the practicing clinician. Despite ongoing educational and preventive services, and despite increasing governmental interdiction, substance use by adolescents continues to be a major national problem. Although present rates of use are lower than the peak rates in the late 1970s, drug use among adolescents nearly doubled in the early 1990s and is a significant cause of morbidity and mortality. Patterns of use continue to evolve. Newer drugs make their way to the streets, and older drugs are rediscovered. Behavioral and environmental factors increase the risk for adolescent substance abuse. Identification of use patterns and familiarity with comorbid behaviors and social risks may help the clinician identify the adolescent at risk.

Use of the Life Optimism Test among adolescents in a clinical setting: a report of reliability testing.

This study examined the consistency and reliability of the Life Optimism Test (LOT) among patients attending an adolescent medicine clinic. The LOT had good internal consistency (Cronbach's alpha ranging from 0.56 at pretest to 0.76 at posttest) and good 1-week test-retest reliability (intraclass correlation = 0.84). These data support use of the LOT to measure optimism among adolescents.

Effectiveness of an inpatient geriatric service in a university hospital.

OBJECTIVE: To assess the effectiveness of an acute interdisciplinary inpatient geriatric service in a university hospital. DESIGN: Prospective randomized control study. SETTING: Large urban university hospital. PATIENTS: 40 consecutive inpatients, randomized for inclusion on the geriatric service (study patients, n = 20) or to continue usual hospital care (control patients, n = 20) from among the geriatric consult population. MAIN OUTCOME MEASURES: Subjects were followed for changes in length of stay, hospital costs, diagnostic testing, pharmacy use, functional status, discharge disposition, and readmission within 30 days after hospitalization. RESULTS: Mean age of patients study 79.2 years (control 73.9 years). Sixty percent of study patients went home and 30% to nursing homes (control 20% home, 65% nursing homes) P = .03. Total length of stay mean 20.3 days study (control 32.7 days), length of stay after randomization mean 7.7 days study (control 11.2 days), mean overall hospital costs $23,906 study (control $45,189), and mean hospital costs after randomization study $4,671 (control $9,404) were not significantly different by F-tests due to wide variability. Laboratory use was reduced with mean 4.4 tests study (control 16.9) P = .01 and mean laboratory costs $263 study (control $828) P = .02. Functional ability improved (scale 1-7) with mean improvement study 0.8 (control 0.3) P = .09. Mean number of medications were lower in the study group by 30% P = .02; mean cost of medications at discharge was reduced with study $38 (control $112); and mean pharmacy charges after randomization decreased $462 study (control $1,268) P = .06. Readmission 30 days after discharge was not significantly different (study 21%, control 33%). CONCLUSIONS: An interdisciplinary acute geriatric service can be cost effective in providing care to elderly patients in a university hospital. It can improve outcomes measured by decreased laboratory and pharmacy usage, improved functional status, and discharge to a lesser level of care without increasing length of stay or early readmission after discharge.

Day case adenoidectomy.

The Audit Commission recommends that more day surgery be undertaken in the Health Service. In a prospective study of 62 children undergoing adenoidectomy one major complication commenced within 2 hours of surgery. Of 15 minor complications, 78% occurred within 6 hours of surgery; they did not herald later major complications. Day adenoidectomy can safely be performed if patients remain in hospital for 6-8 hours post-operatively and then fulfil the following discharge criteria: they are apyrexial, eating, drinking and with no signs of haemorrhage. This will not result in a significant increase in workload for general practitioners. Day adenoidectomy for medically and socially appropriate patients increases our day surgery figures by 74% to 43% of the routine paediatric ENT caseload. There are immediate benefits to patients but the potential economic benefits to the Health Service will not be felt unless these children are treated in a designated Day Case Unit.