Occupational medicine in Canada and Poland in the 1990s

The protocol describes (i) methods for the investigation of neuropeptide catabolism in the central nervous system (CNS), (ii) the identification of the neuropeptidases involved, and (iii) methods for the determination of neuropeptide stability in vitro. These methods are applicable also to study the degradation of peptide hormones by peripheral cells or tissues. To identify peptide degradation products, nanomolar amounts (micromolar concentrations) of peptides are incubated in synthetic media with cell or tissue cultures. Aliquots of the supernatants are withdrawn after different times, peptide fragments separated and fractionated by reversed-phase HPLC, and identified by peptide chemical methods. The peptidases responsible for this degradation can be identified by the use of specific inhibitors listed in the protocol. For receptor binding assays or the study of peptide effects in physiological, nanomolar concentrations the stability of the peptides in an in vitro system should be checked by addition of radiolabeled peptides (femtomolar or nanomolar concentrations) and monitoring the peptide degradation by a procedure analogous to that established for unlabeled peptides. The addition of more or less specific peptidase inhibitors enhances peptide stability in vitro, and thus it can be assured that a given peptide concentration is maintained during biological assays.     

A stable under-gate triode CNT field emitter fabricated via screen printing

A screen printed, under-gate triode with carbon nanotube field emitters is demonstrated. The geometry of the device allows it to be matrix addressable. Fabrication methods including CVD deposition of dielectric and/or nanotubes, wet etching, and photolithography are eliminated by the use of screen printing, effectively lowering the cost of manufacture of the device. Through the use of a charge dissipation layer the emission current is greatly enhanced, and instabilities which plagued previous under-gate devices are eliminated. When a square pulse of 30 µs is applied at a rate of 120 Hz emission currents of > 4 µA/cm² are achieved for gate voltages of < 300 V without any ungated emission. This structure is a prime candidate to be used in a field emission backlight unit (BLU) for an LCD due to its reduced cost of manufacture, stable operation, and local delivery of pulsed light.     

Physical diagnosis skills of physicians in training: a focused assessment

OBJECTIVE: To assess the proficiency of emergency medicine (EM) trainees in the recognition of physical findings pertinent to the care of the critically ill patient. METHODS: Fourteen medical students, 63 internal medicine (IM) residents, and 47 EM residents from three university-affiliated programs in Philadelphia were tested. Proficiency in physical diagnosis was assessed by a multimedia questionnaire targeting findings useful in emergencies or related to diseases frequently encountered in the ED. Attitudes toward diagnosis not based on technology, teaching practices of physical examination during EM training, and self-motivated learning of physical diagnosis also were assessed for all the EM trainees. RESULTS: With the exception of ophthalmology, the EM trainees were never significantly better than the senior students or the IM residents. They were less proficient than the IM residents in cardiology, and not significantly different from the IM residents in all other areas. For no organ system tested, however, did they achieve less than a 42.9% error rate (range: 42.9-72.3%, median = 54.8%). There was no significant improvement in proficiency over the three years of customary EM training. The EM residents who had received supervised teaching in physical diagnosis during training achieved a significantly higher cumulative score. The EM residents attributed great clinical importance to physical diagnosis and wished for more time devoted to its teaching. CONCLUSIONS: These data confirm the recently reported deficiencies of physical diagnosis skills among physicians in training. The results are particularly disturbing because they relate to EM trainees and concern skills useful in the ED. Physical diagnosis should gain more attention in both medical schools and residency programs.     

High-throughput screening: advances in assay technologies

The expression and up-regulation of cell adhesion molecules on a human colonic epithelial cell line HT-29, and the peripheral blood T lymphocyte proliferation responses to bacterial superantigens presented by this cell line were investigated, compared with peripheral blood monocytes. In HT-29 cells, there was constitutive expression of intercellular adhesion molecule-1 (ICAM-1) and lymphocyte function-associated antigen-3 (LFA-3) at a low level, but no constitutive expression of HLA-DR, LFA-1, B7-1 and B7-2 molecules. After stimulation with the supernatants of staphylococcal enterotoxin B (SEB)-stimulated peripheral blood mononuclear cells for 48 h, there was significant up-regulation of HLA-DR and ICAM-1 molecules (both > 90% positive). However, this stimulation had no effect on the expression of LFA-1, B7-1, B7-2 and LFA-3 molecules. In the presence of all tested superantigens SEB, toxic shock syndrome toxin-1, and streptococcal pyogenic exotoxin A, stimulated HT-29 cells caused significant T cell proliferation. When monocytes were used as antigen-presenting cells (APC), the MoAbs against HLA-DR, B7-2 and LFA-3 showed a significant inhibition of SEB-induced T cell proliferation. Anti-ICAM-1 MoAb had no effect on this response. On the other hand, when stimulated HT-29 cells were used as APC, the MoAbs against HLA-DR and ICAM-1 significantly inhibited SEB-induced T cell proliferation. In contrast to monocytes, anti-B7-2 and anti-LFA-3 had no effect on this response. SEB could not induce HT-29 cells to produce IL-8 directly; however, SEB significantly induced the stimulated HT-29 cells to produce IL-8 in the presence of T cells. Thus these data demonstrate that the products of superantigen-stimulated T cell activation can increase the expression of HLA-DR and ICAM-1 molecules on HT-29 cells significantly. Stimulated HT-29 cells can serve as APC to bacterial superantigens. This response is an HLA-DR- and ICAM-1-dependent, but B7-2- and LFA-3-independent process, which was different from professional APC monocytes.     

Characterization of hemolytic and cytotoxic Gallysins: a relationship with arylphorins

The efficacy of injecting antibodies raised against turkey prolactin to prevent the expression of incubation behaviour has been investigated in turkey hens. Medium white turkey hens (n = 15 x 2) were injected three times weekly for 4 consecutive weeks starting on week 5 of egg production. The hens were injected im with a volume of 1 mL per injection for the 1st week and 0.5 mL thereafter, of normal rabbit serum or serum containing antibodies raised against turkey prolactin (Guémené et al, 1994a). None of the 15 passively immunised hens expressed incubation behaviour, whereas, more than half (53%) of the control hens did express it. Plasma prolactin concentrations observed in the two groups presented comparable profiles until week 9 and from week 19 of egg production onward. Differences were, therefore, observed from week 10 until week 17 with the non immunised hens showing higher plasma prolactin concentrations than the immunised ones. This difference was related to the presence of incubating hens in the control group. A higher percentage of non immunised hens disrupted egg production during the course of the study and consequently immunised hens laid more eggs than the control ones. No change in plasma LH and oestradiol concentrations can be related to the immunisation procedure. We conclude that prevention of incubation behaviour can be achieved using passive immunisation against prolactin, prevention which resulted in more egg production under our experimental protocol.     

Molecular cloning and characterization of an invertebrate homologue of a neuropeptide Y receptor

Neuropeptide Y is an abundant and physiologically important peptide in vertebrates having effects on food intake, sexual behaviour, blood pressure and circadian rhythms. Neuropeptide Y homologues have been found in invertebrates, where they are very likely to play similar, important roles. Although five neuropeptide Y-receptor subtypes have been identified in mammals, none has been reported from invertebrates. Here we describe the cloning of a neuropeptide Y-receptor from the brain of the snail Lymnaea stagnalis. The identity of the receptor was deduced by expressing the neuropeptide Y-receptor-encoding cDNA in Chinese Hamster Ovary cells, which were subsequently challenged with size-fractionated Lymnaea brain extracts. An active peptide, selected on the basis of its ability to induce changes in cAMP levels, was purified to homogeneity, analysed by mass spectrometry and amino acid sequence determination, and turned out to be a Lymnaea homologue of neuropeptide Y.     

Survival of Mycobacterium paratuberculosis and preservation of immunoglobulin G in bovine colostrum under experimental conditions simulating pasteurization

OBJECTIVE: To determine whether Mycobacterium paratuberculosis could survive in colostrum after pasteurization. Additionally, this study investigated the effect pasteurization had on IgG concentration in colostrum. ANIMALS: Colostrum samples were collected from cattle (beef and dairy) owned by the state of Ohio. PROCEDURE: Colostrum was divided into aliquots and inoculated with variable concentrations of M paratuberculosis (ATCC No. 19698: 10(4), 10(3), and 10(2) colony-forming units/ml). Half the samples at each concentration were subjected to pasteurization temperatures (63 C) for 30 minutes and the remainder were kept at approximately 20 to 23 C. All samples were incubated (Herrold's egg yolk medium with and without mycobactin J) and observed for growth during the next 16 weeks. Additionally, the IgG concentration of colostrum was determined by radioimmunoassay before and after pasteurization. Samples that coagulated at pasteurization temperatures were mechanically resuspended before measurement of IgG concentration. RESULTS: Growth of M paratuberculosis was retarded but not eliminated by pasteurization. Growth was observed in all unpasteurized samples incubated on Herrold's egg yolk medium with mycobactin J but in only 2 of 18 pasteurized samples similarly cultured. Growth from pasteurized samples appeared 5 to 9 weeks after growth was observed from nonpasteurized samples. Mean colostral IgG concentration was 44.4 g/L in nonpasteurized samples and 37.2 g/L in pasteurized samples, a decrease of 12.3%. High-quality colostrum (> 48 g of IgG/L) had a significantly greater loss of IgG concentration than did colostrum of lesser quality (P = 0.002). CONCLUSIONS: Pasteurization lessened, but did not eliminate, growth of M paratuberculosis from experimentally inoculated colostrum samples. Pasteurization resulted in a significant decrease in colostral IgG concentration but not to an unmanageable level that would preclude the colostrum's use for passive transfer of immunity. CLINICAL RELEVANCE: Colostrum is macrophage rich and may serve as a source of M paratuberculosis infection to calves. Pasteurization of colostrum may lessen the risk of infection, but will not totally eliminate M paratuberculosis.     

Fluvoxamine in the treatment of binge-eating disorder: a multicenter placebo-controlled, double-blind trial

OBJECTIVE: The purpose of this study was to assess the efficacy of fluvoxamine in the treatment of binge-eating disorder. Binge-eating disorder is a newly described eating disorder characterized by recurrent episodes of binge eating but without purging behaviors. Uncontrolled reports have suggested that serotonin selective reuptake inhibitors (SSRIs) may be effective in treating this disorder. METHOD: Eighty-five outpatients with a DSM-IV diagnosis of binge-eating disorder were randomly assigned to receive either fluvoxamine (N=42) or placebo (N=43) in a 9-week, parallel-group, double-blind, flexible dose (50-300 mg) study at three centers. The primary outcome measures were frequency of binge eating, expressed as log ([binges/week]+1), and Clinical Global Impression (CGI) scale ratings. Secondary measures included the level of response (based on the percentage change in frequency of binges), body mass index, and Hamilton Rating Scale for Depression score. Except for the level of response, the outcome measures were analyzed by random regression methods; the treatment-by-time interaction was the measure of treatment effect. RESULTS: Compared with placebo, fluvoxamine was associated with a significantly greater rate of reduction in the frequency of binges, rate of reduction in CGI severity scores, rate of increase in CGI improvement scores, level of response for patients who completed the 9-week study, and rate of reduction in body mass index. There was no significant difference between placebo and fluvoxamine groups in the rate of decrease in Hamilton depression scale scores. A significantly greater proportion of patients receiving fluvoxamine than those receiving placebo discontinued treatment because of an adverse medical event. CONCLUSIONS: In this placebo-controlled trial, fluvoxamine was found to be effective according to most outcome measures in the acute treatment of binge-eating disorder.