Co-detection and discrimination of six human herpesviruses by multiplex PCR-ELAHA.

BACKGROUND: Herpesviruses are a significant cause of human morbidity. Traditional approaches to the identification of these viruses require infectious or at least antigenic virus. Multiplex PCR (mPCR) is capable of simultaneously amplifying a range of targets from a single preparation of nucleic acids and when combined with a suitable detection assay, it is capable of discriminating each of the amplicons. OBJECTIVES: Several methods have been described in the literature, however, they lack one or more significant design features required to suitably control a routinely applied nucleic acid amplification assay. We aimed to design a multiplex herpesvirus PCR that could co-amplify eight human herpesvirus targets plus an internal control (IC) molecule in a single tube. STUDY DESIGN: Primers were designed to target the DNA polymerase genes of each of the human herpesviruses. Synthetic controls were developed to act as templates for the evaluation of assay sensitivity and specificity and for development of an in-house competitive quantitative PCR. Amplicon was discriminated using a simplified enzyme linked amplicon hybridisation assay (ELAHA). RESULTS AND CONCLUSIONS: For routine diagnostic use we reduced the number of herpesviral targets from 8 to 6 in order to maintain adequate clinical sensitivity. The ELAHA proved more sensitive than agarose gel electrophoresis. Additionally, 36 cytomegalovirus positive patients were examined with an in-house quantitative PCR-ELAHA which was developed to confirm that that the mPCR's co-detection limit of 10(2) copy of synthetic template per millilitre was relevant for use in detecting virus from clinical samples. The mPCR-ELAHA was then applied to the screening of 174 patient specimens resulting in a specificity of 98% and a sensitivity of 93%. This preliminary study demonstrated that the mPCR-ELAHA was a complete approach to the detection of herpesviruses from a range of clinical samples and disease states.     

Discoid lupus erythematosus at the site of an intradermal injection of killed staphylococcus aureus

A case of chronic discoid lupus erythematosus is described in which there were recurrent relapses following infections, and at the same time repeated `flare ups' at the site of a delayed hypersensitivity reaction to Staphylococcus aureus. The appearance of the injection site changed from that of a typical `tuberculin reaction' to a patch of discoid lypus erythematosus which was confirmed both histologically and by immunofluorescence.     

A qualitative examination of the implementation of a community–academic coalition

Many recent grant initiatives have mandated coalition building as a key component of their health promotion efforts. Health service leaders are increasingly representing their organizations on coalitions and need to better understand the complexities involved in their creation and management. In this paper we focus on the implementation of a community–academic alliance using an ethnographic approach involving participant observation and in‐depth interviews. Analysis of the data revealed five essential dimensions to be considered in the development of successful community–academic alliances: membership, structure, leadership, communication, and funding. Within each of these areas, facilitators and barriers to successful coalition building were identified. While these areas are not new, obtaining the perspective of coalition members directly adds to our understanding of the member experience in the process of implementing such initiatives. This example of a community–academic collaboration presents one model of how to minimize the distance between research and service and move toward a partnership between these two worlds. © 2004 Wiley Periodicals, Inc. J Comm Psychol 32: 357–374, 2004.     

An investigation into the relationship between salivary cortisol,stress, anxiety and depression

This study examined the relationship between indices of self-reported emotional distress and absolute versus change in cortisol levels. Fifty-four women attending a diagnostic breast clinic completed scales measuring stress, anxiety and depression and provided five saliva samples over the course of a single day for the measurement of cortisol. No significant relationships were evident between absolute cortisol levels and the distress measures. Analysis of the change in cortisol levels revealed a non-linear interaction effect between stress and anxiety and time of day. There was a non-linear relation between time of day and cortisol levels, but the extent of the non-linearity was dependent upon levels of stress and anxiety, not depression. A relationship was apparent between indices of distress and change in cortisol levels, but not absolute levels of the hormone.     

Smoking and Drinking Behavior in Patients with Head and Neck Cancer: Effects of Behavioral Self-Blame and Perceived Control

Patients who continue to use tobacco or alcohol following treatment for head and neck cancers are at greater risk for cancer recurrence and mortality. The present study examined the effects of behavioral self-blame and perceived control over health on smoking and alcohol use in a sample of 55 patients with cancers of the head and neck. Measures of self-blame, perceived control, and depression were administered and an assessment of past and current smoking and drinking behavior was obtained. As anticipated, continued smoking after completion of oncologic treatment was predicted by the interaction of behavior specific self-blame and perceived control. Patients who attributed the cause of their cancer to their past substance use exhibited a lower likelihood of smoking only if they also held the expectancy that their future cancer-related health was contingent on their own behavior. Among patients not holding the belief that cancer recurrence was contingent on their own actions, self-blame was associated with a higher probability of continued smoking. Self-blame and perceived control had no effect on continued alcohol use.     

Functional polymers and sequential copolymers by phase transfer catalysis, 27. Synthesis of aromatic-aliphatic polyformals by polyetherification of 1,4-bis(chloromethoxy)butane with 4,4′-isopropylidenediphenol

The first examples of aromatic-aliphatic polyformals were synthesized from 4,4′-isopropylidenediphenol ( 2 ) and 1,4-bis(chloromethoxy)butane ( 1 ) by a two-phase (aqueous NaOH/toluene), phase transfer catalyzed (PTC) polyetherification. The influence of the reaction conditions upon the polymerization was studied by varying the phase transfer catalyst concentration, NaOH concentration and nucleophile to electrophile ratio. Changes in these parameters and their effects upon the microstructure of these polymers were also examined by ¹H NMR (200 MHz) spectroscopy. The reactivity of 1 has permitted, according to our knowledge, the first examples of interfacial polyetherification. Further, this reactivity has also permitted the homopolymerization of 1 through its hydrolysis byproducts under PTC conditions.     

Different Chinese hamster cell lines express a G1 period for different reasons

Previous studies from our laboratory have shown that the absence of G1 (G1^– condition) in two lines of Chinese hamster cells is dominant over the presence of G1 (G1⁺ condition) in a variety of intraspecific cell hybrids. G1⁺ mutants or variants can be isolated from G1^– cells following mutagenesis and selection. These G1⁺ mutants fall into multiple complementation groups based on their abilities to form G1^– cell hybrids with one another. This is evidence that different mutants have G1 intervals for different reasons, possibly as the result of deficiencies in functions necessary for G1^– cell cycles. In this report we have used cell hybrid analysis to ask whether cells of different, naturally occurring G1⁺ lines of Chinese hamster are able to complement to produce G1^– hybrids. We have found three complementation groups among the four G1⁺ cell lines examined. Therefore, these lines define three different reasons or bases for the existence of a G1 interval. These results lead us to suggest that multiple requirements must be met for these cells to start the S period, but that failure to fulfill only a single and different requirement is responsible for the presence of a G1 interval in any given cell line.     

Thymocyte expression of cathepsin L is essential for NKT cell development

CD1d antigen presentation to natural killer T (NKT) cells expressing the semi-invariant T cell receptor V(alpha)14J(alpha)18 requires CD1d trafficking through endosomal compartments; however, the endosomal events remain undefined. We show that mice lacking the endosomal protease cathepsin L (catL) have greatly reduced numbers of V(alpha)14(+)NK1.1(+) T cells. In addition, catL expression in thymocytes is critical not only for selection of these cells in vivo but also for stimulation of V(alpha)14(+)NK1.1(+) T cells in vitro. CD1d cell-surface expression and intracellular localization appear normal in catL-deficient thymocytes, as does the lysosomal morphology; this implies a specific role for catL in regulating presentation of natural CD1d ligands mediating V(alpha)14(+)NK1.1(+) T cell selection. These data implicate lysosomal proteases as key regulators of not only classical major histocompatibility complex class II antigen presentation but also nonclassical CD1d presentation.     

Linkage analysis of anorexia and bulimia nervosa cohorts using selected behavioral phenotypes as quantitative traits or covariates.

To increase the likelihood of finding genetic variation conferring liability to eating disorders, we measured over 100 attributes thought to be related to liability to eating disorders on affected individuals from multiplex families and two cohorts: one recruited through a proband with anorexia nervosa (AN; AN cohort); the other recruited through a proband with bulimia nervosa (BN; BN cohort). By a multilayer decision process based on expert evaluation and statistical analysis, six traits were selected for linkage analysis (1): obsessionality (OBS), age at menarche (MENAR), and anxiety (ANX) for quantitative trait locus (QTL) linkage analysis; and lifetime minimum body mass index (BMI), concern over mistakes (CM), and food-related obsessions (OBF) for covariate-based linkage analysis. The BN cohort produced the largest linkage signals: for QTL linkage analysis, four suggestive signals: (for MENAR, at 10p13; for ANX, at 1q31.1, 4q35.2, and 8q13.1); for covariate-based linkage analyses, both significant and suggestive linkages (for BMI, one significant [4q21.1] and three suggestive [3p23, 10p13, 5p15.3]; for CM, two significant [16p13.3, 14q21.1] and three suggestive [4p15.33, 8q11.23, 10p11.21]; and for OBF, one significant [14q21.1] and five suggestive [4p16.1, 10p13.1, 8q11.23, 16p13.3, 18p11.31]). Results from the AN cohort were far less compelling: for QTL linkage analysis, two suggestive signals (for OBS at 6q21 and for ANX at 9p21.3); for covariate-based linkage analysis, five suggestive signals (for BMI at 4q13.1, for CM at 11p11.2 and 17q25.1, and for OBF at 17q25.1 and 15q26.2). Overlap between the two cohorts was minimal for substantial linkage signals.