Generation of ß cell-specific human cytotoxic T cells by lentiviral transduction and their survival in immunodeficient human leucocyte antigen-transgenic mice.

Several ß cell antigens recognized by T cells in the non-obese diabetic (NOD) mouse model of type 1 diabetes (T1D) are also T cell targets in the human disease. While numerous antigen-specific therapies prevent diabetes in NOD mice, successful translation of rodent findings to patients has been difficult. A human leucocyte antigen (HLA)-transgenic mouse model incorporating human ß cell-specific T cells might provide a better platform for evaluating antigen-specific therapies. The ability to study such T cells is limited by their low frequency in peripheral blood and the difficulty in obtaining islet-infiltrating T cells from patients. We have worked to overcome this limitation by using lentiviral transduction to 'reprogram' primary human CD8 T cells to express three T cell receptors (TCRs) specific for a peptide derived from the ß cell antigen islet-specific glucose-6-phosphatase catalytic subunit-related protein (IGRP265-273 ) and recognized in the context of the human class I major histocompatibility complex (MHC) molecule HLA-A2. The TCRs bound peptide/MHC multimers with a range of avidities, but all bound with at least 10-fold lower avidity than the anti-viral TCR used for comparison. One exhibited antigenic recognition promiscuity. The ß cell-specific human CD8 T cells generated by lentiviral transduction with one of the TCRs released interferon (IFN)-γ in response to antigen and exhibited cytotoxic activity against peptide-pulsed target cells. The cells engrafted in HLA-A2-transgenic NOD-scid IL2rγ(null) mice and could be detected in the blood, spleen and pancreas up to 5 weeks post-transfer, suggesting the utility of this approach for the evaluation of T cell-modulatory therapies for T1D and other T cell-mediated autoimmune diseases.

From trial to population: a study of a family-based community intervention for childhood overweight implemented at scale.

OBJECTIVES: To assess how outcomes associated with participation in a family-based weight management intervention (MEND 7-13, Mind, Exercise, Nutrition..Do it!) for childhood overweight or obesity implemented at scale in the community vary by child, family, neighbourhood and MEND programme characteristics. METHODS/SUBJECTS: Intervention evaluation using prospective service level data. Families (N=21,132) with overweight children are referred, or self-refer, to MEND. Families (participating child and one parent/carer) attend two sessions/week for 10 weeks (N=13,998; N=9563 with complete data from 1788 programmes across England). Sessions address diet and physical activity through education, skills training and motivational enhancement. MEND was shown to be effective in obese children in a randomised controlled trial (RCT). Outcomes were mean change in body mass index (BMI), age- and sex-standardised BMI (zBMI), self-esteem (Rosenberg scale) and psychological distress (Strengths and Difficulties Questionnaire) after the 10-week programme. Relationships between the outcome and covariates were tested in multilevel models adjusted for the outcome at baseline. RESULTS: After adjustment for covariates, BMI reduced by mean 0.76 kg m(-2) (s.e.=0.021, P<0.0001), zBMI reduced by mean 0.18 (s.e.=0.0038, P<0.0001), self-esteem score increased by 3.53 U (s.e.=0.13, P<0.0001) and psychological distress score decreased by 2.65 U (s.e.=0.31, P<0.0001). Change in outcomes varied by participant, family, neighbourhood and programme factors. Generally, outcomes improved less among children from less advantaged backgrounds and in Asian compared with white children. BMI reduction under service conditions was slightly but not statistically significantly less than in the earlier RCT. CONCLUSIONS: The MEND intervention, when delivered at scale, is associated with improved BMI and psychosocial outcomes on average, but may work less well for some groups of children, and so has the potential to widen inequalities in these outcomes. Such public health interventions should be implemented to achieve sustained impact for all groups.

Chaplains and access to medical records.

This study was initiated by a Pastoral Care Department of a large academic medical center in order to establish hospital chaplaincy policies and procedures. Four basic questions were asked about professional hospital chaplains and record keeping. The results of the survey show that the standard of practice is that chaplains access the medical record, enter notes in the record, have access to the electronic medical record, and that no special credentialing beyond Clinical Pastoral Education (CPE) is required for chaplains to have this access.

Developing a Strategic Plan for Transitioning to Healthcare Knowledge Services Centers (HKSCs).

Facing a negative trend in the form of downsizing, layoffs, and closures, a small committee of hospital librarians in New England was formed in 2004 to provide library advocacy. Between 2008 and 2010, 23 hospital libraries closed in New England. In 2010, the committee shifted its focus from advocacy to a platform for change. This resulted in the creation of the Healthcare Knowledge Services Center (HKSC) Template. The Template is the basis for a 3-phased, 5-year strategic plan to establish several regional pilots, transitioning traditional hospital libraries to healthcare knowledge services centers. This article focuses on Phase One of the strategic plan, Development.

Lentivectors encoding immunosuppressive proteins genetically engineer pancreatic beta-cells to correct diabetes in allogeneic mice.

The effectiveness of genetic engineering with lentivectors to protect transplanted cells from allogeneic rejection was examined using, as a model, type 1 diabetes treatment with beta-cell transplantation, whose widespread use has been limited by the requirement for sustained immunosuppressive treatment to prevent graft rejection. We examined whether lentivectors expressing select immunosuppressive proteins encoded by the adenoviral genome early region 3 (AdE3) would protect transplanted beta-cells from an alloimmune attack. The insulin-producing beta-cell line beta TC-tet (C3HeB/FeJ-derived) was transduced with lentiviruses encoding the AdE3 proteins gp19K and RID alpha/beta. The efficiency of lentiviral transduction of beta TC-tet cells exceeded 85%. Lentivector expression of gp19K decreased surface class I major histocompatibility complex expression by over 90%, whereas RID alpha/beta expression inhibited cytokine-induced Fas upregulation by over 75%. beta TC-tet cells transduced with gp19K and RID alpha/beta lentivectors, but not with a control lentivector, provided prolonged correction of hyperglycemia after transplantation into diabetic BALB/c severe combined immunodeficient mice reconstituted with allogeneic immune effector cells or into diabetic allogeneic BALB/c mice. Thus, genetic engineering of beta-cells using gp19K- and RID alpha/beta-expressing lentiviral vectors may provide an alternative that has the potential to eliminate or reduce treatment with the potent immunosuppressive agents necessary at present for prolonged engraftment with transplanted islets.

Depression, abuse and its relationship to interstitial cystitis.

This study evaluated the prevalence of depression, sexual abuse, and physical abuse among women diagnosed with interstitial cystitis (IC). One hundred forty-one subjects completed the validated Beck's Depression Inventory II Questionnaire (BDI-II) and the validated Drossman Abuse Questionnaire. Ninety-seven (69%) subjects scored 14 or higher on the BDI-II, corresponding to depression. When compared to the US prevalence of 9%, this was significantly higher. Fifty-one subjects (36%) reported sexual abuse which is higher than the US average. The prevalence of childhood sexual abuse in the sample was not significantly different than the US average. The prevalence of physical abuse in the sample was not statistically different than the US average. Women with IC appear to have a higher prevalence of depression and sexual abuse than the general population. Women with IC should be screened for depression and abuse and referred to a mental health expert as necessary for treatment.

Evaluation of pain and disability in plug repair with the aid of a personal digital assistant.

The purpose of this paper is to illustrate the method of real-time data collection using a hand-held personal digital assistant (PDA) in the operating suite, hospital, and office. The technique for the placement of a round Atrium ProLite self-forming, layered polypropylene plug in 155 hernia repairs is described. The study measured postoperative pain by the number of pills used (6.6) and categorizes patient return to work according to sedentary, manual, and standing with an average return to work time of 15.6 days for unilateral repair and 20.4 days for bilateral repair. Return to normal daily activity was within 22.3 days for unilateral- and 28.5 days for bilateral repair.

Three strategies for changing attributions about severe mental illness.

The effects of three strategies for changing stigmatizing attitudes--education (which replaces myths about mental illness with accurate conceptions), contact (which challenges public attitudes about mental illness through direct interactions with persons who have these disorders), and protest (which seeks to suppress stigmatizing attitudes about mental illness)--were examined on attributions about schizophrenia and other severe mental illnesses. One hundred and fifty-two students at a community college were randomly assigned to one of the three strategies or a control condition. They completed a questionnaire about attributions toward six groups--depression, psychosis, cocaine addiction, mental retardation, cancer, and AIDS--prior to and after completing the assigned condition. As expected, results showed that education had no effect on attributions about physical disabilities but led to improved attributions in all four psychiatric groups. Contact produced positive changes that exceeded education effects in attributions about targeted psychiatric disabilities: depression and psychosis. Protest yielded no significant changes in attributions about any group. This study also examined the effects of these strategies on processing information about mental illness.

Stereotactic breast biopsy of nonpalpable lesions: determinants of ductal carcinoma in situ underestimation rates.

PURPOSE: To measure the effect of biopsy device, probe size, mammographic lesion type, lesion size, and number of samples obtained per lesion on the ductal carcinoma in situ (DCIS) underestimation rate. MATERIALS AND METHODS: Nonpalpable breast lesions at 16 institutions received a histologic diagnosis of DCIS after 14-gauge automated large-core biopsy in 373 lesions and after 14- or 11-gauge directional vacuum-assisted biopsy in 953 lesions. The presence of histopathologic invasive carcinoma was noted at subsequent surgical biopsy. RESULTS: By performing the chi(2) test, independent significant DCIS underestimation rates by biopsy device were 20.4% (76 of 373) of lesions diagnosed at large-core biopsy and 11.2% (107 of 953) of lesions diagnosed at vacuum-assisted biopsy (P <.001); by lesion type, 24.3% (35 of 144) of masses and 12.5% (148 of 1,182) of microcalcifications (P <.001); and by number of specimens per lesion, 17.5% (88 of 502) with 10 or fewer specimens and 11.5% (92 of 799) with greater than 10 (P <.02). DCIS underestimations increased with lesion size. CONCLUSION: DCIS underestimations were 1.9 times more frequent with masses than with calcifications, 1.8 times more frequent with large-core biopsy than with vacuum-assisted biopsy, and 1.5 times more frequent with 10 or fewer specimens per lesion than with more than 10 specimens per lesion.

A university experience using mesh in inguinal hernia repair.

In a university program, surgical residents are trained to perform classic inguinal hernia repair. The necessity of this approach is discussed. In this study, polypropylene mesh was added to the repair in order to familiarize the resident with the properties and qualities of mesh. Three hundred seventy-one consecutive herniorrhaphies were performed using a precut mesh in patients weighing between 110 lbs and 285 lbs. All patients were seen postoperatively, and 120 were seen at 1 year. One recurrence and no major complications were noted. Deficient local collagen and properties of mesh prosthetics are discussed as part of the surgical resident curriculum.

The relationship of acculturation and gender to attitudes toward counseling in Italian and Greek American college students.

This exploratory study examined attitudes toward professional psychological services and help provider characteristics among 232 self-identified Italian and Greek American college students in 3 Northeast colleges. Regarding general attitudes toward psychological services in the Italian American sample, women had a greater recognition of personal need for help and higher confidence in the ability of mental health professionals to meet these needs than did men. With regard to preferred counselor demographic characteristics, regardless of gender, lower acculturated Italian American students had a stronger preference for seeing an ethnically similar counselor. With the Greek American sample, there was an interaction effect between acculturation level and gender on attitudes toward services. Among the higher acculturated Greek students, women were more open regarding their personal concerns than men; however, within the lower acculturated cohort, no gender differences were found. With regard to counselor characteristic variables, and similar to Italian Americans, regardless of gender, lower acculturated Greek Americans had a stronger preference for seeing an ethnically similar counselor to discuss a personal problem. Limitations and implications for further research are noted.

Social stories, written text cues, and video feedback: effects on social communication of children with autism.

This study investigated the effects of written text and pictorial cuing with supplemental video feedback on the social communication of 5 students with autism and social deficits. Two peers without disabilities participated as social partners with each child with autism to form five triads. Treatment was implemented twice per week and consisted of 10 min of systematic instruction using visual stimuli, 10 min of social interaction, and 10 min of self-evaluation using video feedback. Results showed increases in targeted social communication skills when the treatment was implemented. Some generalized treatment effects were observed across untrained social behaviors, and 1 participant generalized improvements within the classroom. In addition, naive judges reported perceived improvements in the quality of reciprocal interactions. These findings support recommendations for using visually cued instruction to guide the social language development of young children with autism as they interact with peers without disabilities.

A multilevel model framework for meta-analysis of clinical trials with binary outcomes.

In this paper we explore the potential of multilevel models for meta-analysis of trials with binary outcomes for both summary data, such as log-odds ratios, and individual patient data. Conventional fixed effect and random effects models are put into a multilevel model framework, which provides maximum likelihood or restricted maximum likelihood estimation. To exemplify the methods, we use the results from 22 trials to prevent respiratory tract infections; we also make comparisons with a second example data set comprising fewer trials. Within summary data methods, confidence intervals for the overall treatment effect and for the between-trial variance may be derived from likelihood based methods or a parametric bootstrap as well as from Wald methods; the bootstrap intervals are preferred because they relax the assumptions required by the other two methods. When modelling individual patient data, a bias corrected bootstrap may be used to provide unbiased estimation and correctly located confidence intervals; this method is particularly valuable for the between-trial variance. The trial effects may be modelled as either fixed or random within individual data models, and we discuss the corresponding assumptions and implications. If random trial effects are used, the covariance between these and the random treatment effects should be included; the resulting model is equivalent to a bivariate approach to meta-analysis. Having implemented these techniques, the flexibility of multilevel modelling may be exploited in facilitating extensions to standard meta-analysis methods.

Multivariate spatial models for event data.

This paper describes how estimates made for event rates in small areas may be enhanced through spatial modelling of the data - taking the geographical location of each area into account - and through the addition of further information from each area. In particular we consider the use of spatial models to predict more than one outcome simultaneously. This is done by writing the spatial model as a multi-level model and subsequently enhancing this to encompass a multivariate data structure; estimates are obtained using iterative generalized least squares in the software package MLwiN. The example given considers mortality due to two causes--neoplasms and circulatory disease--in 143 postcode sectors in Greater Glasgow Health Board, Scotland. In addition, a measure of socio-economic deprivation is available for each area. Correlations between causes within areas, between areas within causes and between areas and causes are quantified, as are the relative contributions of the heterogeneous and spatial parts of the model. The results suggest a tendency for there to be pockets with high mortality rates due to neoplasms, whilst mortality due to circulatory disease follows a much smoother pattern. After taking deprivation into account, the spatial component accounts for just 19 per cent of the variation in the mortality due to neoplasms in Greater Glasgow but 66 per cent of the mortality due to circulatory disease.

Efficient gene transfer to hematopoietic progenitor cells using SV40-derived vectors.

We used recombinant SV40 (rSV40)-derived vectors to deliver transgenes to human and simian hematopoietic progenitor cells in culture, and in vivo after transduction ex vivo. rSV40 are highly efficient vectors that are made in very high titers. They infect almost all cells, whether resting or dividing. Two rSV40s were used: SV(HBS), carrying hepatitis B surface antigen as a marker; and SV(Aw) carrying IN#33, a single chain Fv antibody against HIV-1 integrase. CD34+ cells derived from human fetal bone marrow (HFBM) and rhesus macaque bone marrow were transduced once with SV(HBS) without selection. On average 60% of colonies derived from transduced CD34+ cells carried and expressed HBsAg, as assessed by PCR and immunochemistry. Transgene carriage persisted following differentiation of transduced rhesus CD34+ cells into T lymphocytes. In an effort to increase the percentage of gene-marked cells, three sequential treatments of CD34+ cells were done using SV(Aw), without selection. Two weeks later, >95% of colonies expressed IN#33. Unselected SV(Aw)-transduced CD34+ cells from HFBM were transplanted into sublethally irradiated SCID mice. Bone marrow harvested 3 months later showed that >50% of bone marrow cells expressed IN#33. This is comparable with the percentage of human cells in these animals' bone marrow as judged by immunostaining for human CD45. The stability and longevity of transduction in this setting suggests that rSV40 vectors integrate into the cellular genome. This possibility was supported by finding that PCR of genomic DNA using primer pairs with one cellular and one viral primer yielded PCR products only in transduced, but not control, cells. These PCR products hybridized with an SV40 DNA fragment. Thus, rSV40 vectors transduce normal human and primate bone marrow progenitor cells effectively without selection, and maintain transgene expression in vivo following reimplantation. Such high efficiency transduction may be useful in treating diseases of CD34+ cells and their derivatives.

The utility of thyroid nuclear imaging and other studies in the detection and treatment of underlying thyroid abnormalities in patients with endogenous subclinical thyrotoxicosis.

PURPOSE: Endogenous subclinical thyrotoxicosis is diagnosed when a patient who is not taking exogenous thyroid hormone has a suppressed level of thyroid-stimulating hormone with normal levels of the free thyroid hormones thyroxine and triiodothyronine and other known causes of a suppressed thyroid-stimulating hormone level have been excluded. Although such a condition is caused by underlying thyroid disease, the specific nature and relative prevalence of these disorders and the utility of nuclear imaging and other studies in their detection remains unclear. PATIENTS AND METHODS: The authors performed a retrospective study of 50 patients with endogenous subclinical thyrotoxicosis. The results of the history and physical examination, thyroid nuclear scan, radioactive iodine uptake measurement, and thyroid antibody studies were reviewed. The results of the nuclear imaging and thyroid antibody studies were combined in an attempt to establish an underlying diagnosis for each patient. RESULTS: The thyroid nuclear imaging and antibody studies were used to establish a specific thyroid disorder in most of the patients (n = 39). These disorders included most commonly toxic multinodular goiter, various forms of autoimmune thyroid disease, and solitary toxic adenoma. A specific diagnosis was not determined in 11 patients. Therapy with I-131 radioactive iodine was administered to 14 of these patients, 13 of whom subsequently achieved a normal thyroid-stimulating hormone level. CONCLUSIONS: Most patients with endogenous subclinical thyrotoxicosis have underlying thyroid abnormalities that can be determined by nuclear imaging and, in selected cases, thyroid antibody studies.