Compartmentalization of Th1/Th2 cytokine responses to experimental Yersinia pseudotuberculosis infection in cats

Specific pathogen-free cats were inoculated subcutaneously into the drainage areas of the left auricular and popliteal lymph nodes with living Yersinia pseudotuberculosis. Inflammation was evident at the inoculation sites and the regional lymph nodes were palpably enlarged at 48 h post-infection. Lymph node enlargement was due to marked paracortical lymphoid hyperplasia and variable neutrophil infiltrates. Yersinia was cultured from the regional lymph nodes and/or spleens of three of the six cats, indicating systemic spread of bacteria. Specific T-helper 1 and 2 (Th1, Th2) cell-associated cytokine mRNA levels were compared in regional lymph nodes, peripheral blood mononuclear cells (PBMC) and spleen at 48 h post-inoculation. Relative to unstimulated control tissues, there was a significant increase in TNF-alpha, IFN-gamma, IL-12, and IL-10 mRNAs in spleen with down-regulation of IL-4. Significant up-regulation of TNF-alpha and down-regulation of IL-4 were also observed in PBMC. Paradoxically, 48 h stimulated lymph nodes showed only minimal differences in cytokine mRNA expression when compared to lymph nodes from mock-inoculated control animals or unchallenged contralateral lymph nodes from the same animal. This study demonstrated that cats, like mice, respond to an intracellular pathogen such as Y pseudotuberculosis with a predominantly Th1-type immune response. The cytokine responses in regional lymph nodes and spleen were asynchronous, while cytokine stimulation in cells of the spleen was mirrored by PBMC.     

Amphetamines in hair by enzyme-linked immunosorbent assay

Human hair was collected from the occipital crown region of the head from several subjects; these hair samples were presumptively positive for amphetamines by a previously evaluated immunoassay. Hair was washed briefly with methanol to remove external contamination, then extracted with hot methanol for 2 h to recover the drugs. The extracts were evaporated to dryness, reconstituted in buffer, and analyzed using a new enzyme-linked immunosorbent assay (ELISA) technique adapted for the detection of amphetamines in hair. Gas chromatography-mass spectrometry was used as the reference technique. Cross-reactivity of several related compounds was evaluated by equating the inverse of the ligand concentration at 50% antibody binding to the affinity constant for each compound. The ratio of a compound's affinity constant to that for d-methamphetamine was used to derive percent crossreactivity. These experiments yielded values of 30.8% for d-amphetamine, 7.4% for I-methamphetamine, 4.3% for phentermine, 2.9% for I-amphetamine, and <1% for ephedrine, methylenedioxyamphetamine, and methylenedioxymethamphetamine. Cross-reactivity of unrelated compounds was found to be non-existent. The optimum cutoff concentration was determined by receiver operating characteristic curve analysis to be 300 pg/mg and the observed limit of detection was 60 pg/mg. Intra-assay precision at 300 pg/mg was 3.3% (coefficient of variation, CV), and the interassay CV was 10.5%. The sensitivity and specificity of the method were 83% and 92%, respectively.     

Patient choice in diabetes education curriculum. Nutritional versus standard content for type 2 diabetes

The purpose of this study was to determine whether aging alters neuronal uptake of norepinephrine (NE) in the rat heart and if dietary restriction influenced the effect of age on this system. Cardiac synaptosomes were prepared from 6-, 12- and 24-month-old male F344 rats fed ad libitum (AL) or a diet restricted (DR) to 60% of AL intake. Cardiac synaptosomes were incubated with 50, 100, 200, or 400 nM [3H]NE for 10 min at 37 degrees C with and without desmethylimipramine (DMI), a selective neuronal-uptake blocker. DMI-sensitive [3H]NE uptake was calculated as the difference between samples with and without DMI. NE uptake was adjusted for the number of cardiac synaptosomes in each sample by dividing by the endogenous NE content in each sample. The Vmax for uptake ([3H]NE/min/ng NE) declined significantly between 6 and 12 months in AL rats and between 12 and 24 months in DR rats. Km was not significantly different between age or diet groups. The change in Vmax with age suggests that the number of NE transporters per synaptosome may decline with age and that DR delays this effect of age. There were no differences in the sensitivity to DMI between age or diet groups.     

Reported changes in sexual behavior in response to AIDS in the United States

BACKGROUND: The AIDS epidemic has profoundly affected public health policy and everyday choices of individuals. Understanding the self-protective strategies adopted by the general population is crucial to designing appropriate and more effective primary AIDS prevention strategies. METHODS: A probability sample of the U.S. adult population in 1992, the National Health and Social Life Survey (n = 3,159), addressed detailed aspects of sexual behavior. How respondents changed their sexual behavior due to AIDS was recorded verbatim and categorized for analysis. RESULTS: Twenty-nine percent of U.S. adults reported some sexual behavior change, primarily, reducing numbers of partners (12%), using condoms more frequently (9%), selecting partners more carefully (7%), changing attitude toward sex (4%), and abstinence (3%). Individuals most at risk show the greatest propensity to change although the actual risk-reduction strategies and the meanings assigned to those strategies vary considerably. Strategies are not all compatible with each other nor equally likely to be adopted by individuals with different lifestyles. CONCLUSIONS: Self-protective strategies to avoid AIDS are widespread, but primary prevention efforts should be targeted to particular circumstances and the self-protective propensities of each individual.     

Comparison of measured sleeping metabolic rate and predicted basal metabolic rate in the first year of life

In infants, sleeping metabolic rate (SMR) is used as a proxy for basal metabolic rate (BMR). BMR can be predicted from anthropometry using published equations. Our study was intended to evaluate the ability of these equations to predict measured SMR in infants aged 6 weeks to 12 months. SMR was measured in a mixed longitudinal study using the Douglas bag technique (n = 105). Measured SMR values were compared with BMR predicted from weight (BMR-1) or weight and length (BMR-2). These equations were not successful in predicting SMR in this age group. Percentage error of predicted BMR was related to infant weight (BMR-1: r = 0.26; p < 0.005; BMR-2: r = 0.18; p < 0.06). Alternative logarithmic equations were derived from this study (R = 0.84-0.87; SEE = 0.159-0.168). We conclude that the new equations, relating to contemporary infants, are more suitable but actual measurements remain preferred.     

Molecular cloning, sequencing, and heterologous expression of the vaoA gene from Penicillium simplicissimum CBS 170.90 encoding vanillyl-alcohol oxidase

The cDNA encoding vanillyl-alcohol oxidase (EC 1.1.3.7) was selected from a cDNA library constructed from mRNA isolated from Penicillium simplicissimum CBS 170.90 grown on veratryl alcohol by immunochemical screening. The vaoA-cDNA nucleotide sequence revealed an open reading frame of 1680 base pairs encoding a 560-amino acid protein with a deduced mass of 62,915 Da excluding the covalently bound FAD. The deduced primary structure shares 31% sequence identity with the 8alpha-(O-tyrosyl)-FAD containing subunit of the bacterial flavocytochrome p-cresol methyl hydroxylase. The vaoA gene was isolated from a P. simplicissimum genomic library constructed in lambdaEMBL3 using the vaoA-cDNA as a probe. Comparison of the nucleotide sequence of the vaoA gene with the cDNA nucleotide sequence demonstrated that the gene is interrupted by five short introns. Aspergillus niger NW156 prtF pyrA leuA cspA transformed with the pyrA containing plasmid and a plasmid harboring the complete vaoA gene including the promoter and terminator was able to produce vaoA mRNA and active vanillyl-alcohol oxidase when grown on veratryl alcohol and anisyl alcohol. A similar induction of the vaoA gene was found for P. simplicissimum, indicating that similar regulatory systems are involved in the induction of the vaoA gene in these fungi. Introduction of a consensus ribosome binding site, AGAAGGAG, in the vaoA-cDNA resulted in elevated expression levels of active vanillyl-alcohol oxidase from the lac promoter in Escherichia coli TG2. The catalytic and spectral properties of the purified recombinant enzyme were indistinguishable from the native enzyme.     

Long-term follow-up of an EORTC randomized prospective trial comparing intravesical bacille Calmette-Guérin-RIVM and mitomycin C in superficial bladder cancer. EORTC GU Group and the Dutch South East Cooperative Urological Group. European Organisation for Research and Treatment of Cancer Genito-Urinary Tract Cancer Collaborative Group

Posterior laryngeal clefts (PLCs) are described in the literature as rare laryngeal abnormalities. The authors believe type I clefts are much more common than previously reported. In two busy pediatric tertiary care centers, such clefts are the second most common congenital laryngeal finding at rigid endoscopy, second only to laryngomalacia. PLCs frequently present with symptomatology that can be attributed to other common disease processes and are often undiagnosed unless the surgeon maintains a high index of suspicion and specifically examines the posterior glottis by palpation during microlaryngoscopy. This report presents a series of 41 patients with type I PLCs, reviews their subtle and often confusing presenting signs and symptoms, and describes a simple yet reliable method of diagnosis.     

Aboriginal perspectives of diabetes in a remote community in the Northern Territory

This study explores the knowledge and beliefs of diabetes in a group of Aboriginal people from a remote community in the Northern Territory. Information was gathered from participants through a combination of group discussions, semi-structured interviews and informal conversations. The four themes of explanation of diabetes were: worry, food, family and infections. The most common means of preventing or treating diabetes were stopping worry and changing one's diet. Significantly, few of the participants believed that medication was effective, and weight loss and exercise were not mentioned. The results highlight the miscommunication about diabetes that has occurred in this population. For effective communication and management of diabetes, health information must incorporate the contemporary health beliefs of the clients.