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Correlation of plasma corticosterone levels with running activity in the blinded rat.

Blinded female reats were placed in running-wheel cages to monitor the phase of their activity cycle. After approximately a month of adaptation to the wheels, jugular vein blood samples were withdrawn at the beginning and end of the running phase of activity and analyzed for plasma corticosterone. By the time blood samples were obtained, the blined rats' activity cycles had desynchronized from the lighting cycle of the room in an apparently free-running fashion, and were out of phase with each other. The corticosterone levels were high at the beginning of the running phase and low at the end: begin run corticosterone was 34.5 +/- 13, end run corticosteronewas 14.2 +/- 10 (mean +/-SD in mug/100 ml of plasma; t = 7.93, df = 82, P less than 0.001). It was concluded that blinded rats do have an adrenocortical rhythm and that it is in phase with the activity cycle of each individual rat.

Adrenal Glands↗

Beta-endorphin, but not oxytocin, substance P or vasoactive-intestinal polypeptide, contributes to progesterone-induced prolactin secretion in monkeys.

Progesterone (P) stimulates prolactin secretion through a neural mechanism in estrogen (E)-primed female monkeys. Several peptides, including beta-endorphin (BE), oxytocin (OT), substance P (SP) and vasoactive intestinal polypeptide (VIP) are potential prolactin stimulatory factors and could mediate the effect of P. We hypothesized that the antagonism of a pivotal peptidergic neural system would block P-induced prolactin secretion and that the function of a pivotal peptidergic system would be altered by changes in gonadal steroid concentrations. Therefore it was of interest (1) to examine the effect of infusion of antagonists to these peptides on P-induced prolactin secretion, and (2) to determine BE, OT, SP and VIP levels in the hypothalamus of monkeys of various reproductive states. For the antagonist studies, female monkeys (n = 8) were spayed, adapted to a vest and tether remote sampling system and catheterized prior to antagonist challenges. E-primed monkeys received P injections 48 h prior to antagonist administration. Prolactin increased within 36-48 h of P injection. All antagonist challenges were administered in varying doses during the P-induced prolactin elevation and blood samples were collected every 10 min for prolactin determinations. The opiate antagonist, naloxone (n = 5), reduced serum prolactin in a dose-related manner with a mean IC50 of 1.5 +/- 0.6 micrograms/kg/min. The OT (n = 4), SP (n = 4) or VIP (n = 4) antagonists did not reduce serum prolactin in a dose-related manner. We previously reported that the hypothalamic content of OT is increased by ovarian hormones. To determine whether the hypothalamic content of BE, SP or VIP was related to gonadal status, the peptide levels in 4 hypothalamic regions of monkeys in various physiological states were measured. BE (ng/mg protein) in the medial basal hypothalamus (MBH) was significantly greater in adult females (17.7 +/- 6.9; n = 6) as compared to spayed females (0.6 +/- 0.2; n = 3) and juvenile females (1.8 +/- 1.1; n = 3). Hypothalamic content of SP in the preoptic area and mammillary bodies, but not the MBH, was significantly greater in gonadal intact females than spayed females. VIP content (pg/mg protein) was not significantly different between adult, spayed and juvenile females nor between adult and juvenile males in any hypothalamic area. Taken together these results support a pivotal role for BE in the neural regulation of P-induced prolactin secretion. The involvement of OT, SP, and VIP in a specific manner at the pituitary level is not indicated.

Animals↗

RU 486 blocks and fluoxetine augments progesterone-induced prolactin secretion in monkeys.

Progesterone (P) stimulates prolactin secretion through an unknown neural mechanism in estrogen (E)-primed female monkeys. Serotonin also stimulates prolactin secretion and this laboratory demonstrated that E induces nuclear progestin receptors (PR) in serotonin neurons. Thus, PR in serotonin neurons could transduce the action of P on prolactin secretion. Studies were performed to determine (1) whether blocking nuclear PR would block P-induced prolactin secretion and conversely; (2) whether increasing serotonin concentrations in the synapse would augment P-induced prolactin secretion. In both studies, female monkeys were spayed, adapted to a vest and tether remote sampling system and catheterized prior to experiments. Monkeys received 2 E-filled silastic implants (3.0 cm) 1-3 weeks prior to study. P (20 mg) in corn oil was injected (s.c.) to transiently increase prolactin secretion. In both studies, each monkey served as its own control. To block nuclear PR and not membrane PR, RU 486 (2 mg/kg, i.m.) or ethanol (control) was administered with the P injection. Relative to the P injection, blood samples were taken twice daily from -30 to +24 h, then every 4 h from +36 to +48 h and once at +65 h. To increase serotonin in the synapse, the serotonin reuptake inhibitor, fluoxetine (5 mg/day, i.v.), was infused for 4 weeks. P was injected during the week of vehicle infusion and during the last week of fluoxetine infusion. Blood samples were obtained twice daily prior to and following P treatment. Prolactin, E, P and RU 486 concentrations were determined by RIA. RU 486 completely blocked the P-induced prolactin surge (n = 3). In addition, fluoxetine significantly increased prolactin secretion during the P-induced prolactin peak compared to equal time points during saline infusion (n = 5). These data indicate that P induces prolactin via a genomic mechanism and not through a membrane action. The data also support a pivotal role for serotonin in the neural regulation of P-induced prolactin secretion.

Animals↗

A single subtype of avian pneumovirus circulates among Minnesota turkey flocks.

The recent emergence of avian pneumovirus (APV) infection among US turkey flocks has resulted in a major economic threat to the turkey industry. In order to elucidate the molecular epidemiology of APV, comparative sequence analysis of the fusion (F) protein gene of APV was performed for 3 cell culture-adapted isolates and 10 APV positive clinical samples recovered from US turkey flocks. Relatively modest levels of nucleotide and amino acid sequence divergence were identified, suggesting the prevalence of a single lineage of APV among US turkey flocks. Additionally, numerous polymorphisms were identified that were only represented in the clinical samples but not in the in vitro propagated isolates of APV. Phylogenetic analyses confirm that the subtype of APV circulating in the upper Midwestern United States is evolutionarily related to, but distinct from, European APV subgroups A and B. Overall, the results of the present investigation suggest that there has been only a single recent introduction of APV into US turkey populations in the upper Midwestern United States.

Amino Acid Sequence↗

Luteal phase deficiency: abnormal gonadotropin and progesterone secretion patterns.

Luteal phase deficiency (LPD) is a reproductive disorder associated with infertility and spontaneous abortion. This study was undertaken to determine whether LPD might be related to an abnormal pattern of gonadotropin secretion. We tested this hypothesis by evaluating the pattern of pulsatile LH secretion in both the follicular and luteal phases of the menstrual cycle in normal women (n = 21) and women with LPD (n = 20), which was diagnosed on the basis of two out of phase endometrial biopsies. In addition, we sought to determine whether changes in progesterone (P) pulse patterns could account for the decrease in average serum P levels in women with LPD. To this end, we examined the pulse patterns of P and compared these patterns between normal women and those with LPD. Frequent blood sampling was performed in both groups to determine their respective hormone secretion patterns. In the follicular phase, blood samples were obtained every 10 min for 12 h; in the luteal phase the samples were obtained every 10 min for 12 h; in the luteal LH, FSH, and P were assayed in each sample. Pulse detection was performed by an adaptive threshold method of pulse analysis. The LH pulse frequency was significantly higher in the women with LPD than in the normal women in the early follicular phase [P less than 0.05; LPD, 12.8 +/- 1.4 (+/- SE); normal, 8.2 +/- 0.7 pulses/12 h]. LH pulse frequency was similar in the early and late follicular phases in the women with LPD, whereas it was higher in the late follicular phase in normal women. Mean serum FSH levels were not different between groups in both the early and late follicular phases. In the luteal phase the P pulse amplitude and mean serum P level were significantly lower in the LPD group than in the normal women (P less than 0.01). We conclude that 1) a too rapid LH pulse pattern in the early follicular phase may lead to inadequate LH support of the corpus luteum and become manifest as LPD; 2) the mechanism for inadequate P secretion in LPD is decreased P pulse amplitude; 3) the finding of similar serum FSH levels in the two groups in both the early and late follicular phases did not support compromised folliculogenesis as an etiological factor for LPD.

Abortion, Habitual↗

An immunogold cell labelling method for rat T lymphocytes.

A method using monoclonal antibodies in conjunction with an immunogold procedure was adapted for labelling T lymphocytes in blood samples obtained from male Wistar rats. The monoclonal antibodies W3/25, MRC/OX8 and W3/13 were used. Changes in the percentages of positively labelled cells were observed in rats dosed with the immunosuppressant cyclosporin.

Animals↗

Simplified diagnosis of malaria infection: GFM/PCR/ELISA a simplified nucleic acid amplification technique by PCR/ELISA.

We report an adaptation of a technique for the blood sample collection (GFM) as well as for the extraction and amplification of Plasmodium DNA for the diagnosis of malaria infection by the PCR/ELISA. The method of blood sample collection requires less expertise and saves both time and money, thus reducing the cost by more than half. The material is also suitable for genetic analysis in either fresh or stored specimens prepared by this method.

Animals↗

Financial risk relationships and adoption of management strategies in physician groups for self-administered injectable drugs.

OBJECTIVE: To consider the extent, nature, and range of risk arrangements between physician groups and health maintenance organizations (HMOs) for self-administered injectable (SAI) drugs; to examine types and frequencies of SAI drug-use management strategies adopted by physician groups; and to explore the relationship between locus and level of financial risk for SAIs and physician group strategy adoption. METHODS: We used a multiple case-study design to select physician groups and their health maintenance organization (HMO) contractual partners in 4 markets in the United States (Northwest, Northeast, Midwest, Southwest). Physician groups in these markets were chosen based on size (e50 physicians) and experience with drug risk (e1 year). Physician groups were asked to identify their 3 major HMO contractual partners in each market. Telephone interviews were conducted from January 2000 to June 2001, with the resulting purposive sample of 37 individuals representing 20 physician groups. RESULTS: We found that the level and locus of SAI financial risk were related to the adoption of management strategies. Physician groups with higher financial risk for SAIs adopted more strategies than lower-risk groups. Groups with SAI financial risk in the medical services capitation (MSC) adopted 9.2 strategies per group. In contrast, groups with SAI financial risk in the pharmacy-risk budget (PRB) averaged 1.5 strategies per group. Groups with SAI financial risk in both the MSC and PRB fell in-between, averaging 4.5 strategies per group. The most frequently adopted strategy was designing evidenced-based therapeutic guidelines, i.e., protocols based on evidence from the peer-reviewed literature used to guide physicians in the treatment of typically chronic conditions (9 groups, 45% of sample). The second most common strategy involved adapting the existing utilization management system to process SAIs (7 groups, 35%) and the establishment of office procedures for internal authorization (5 groups, 25%). The least frequently used strategies were determining amount paid to out-of-group physician providers (1 group, 5%) and hiring personnel (e.g., pharmacists) in claims or utilization management departments to implement and manage SAI programs (1 group, 5%). We also identified potential factors that increased the likelihood of strategy adoption and that could slow the rate of SAI cost increases. CONCLUSION: Our findings suggest that adoption of SAI drug-use management strategies may be more likely to occur when there is a minimum level of risk for SAI drug costs. Likewise, both the adoption of strategies and the opportunity to slow the rate of SAI cost increases may be more likely to occur when 3 additional factors are present: a contractual environment conducive to controlling SAI drug costs, the ability to implement SAI drug-use management strategies, and power in negotiations with drug manufacturers to reduce SAI prices. A sustainable and affordable SAI financial risk management program maximizing these factors while minimizing the financial burden for patients will require collaboration among all stakeholders, payers, providers, drug manufacturers, and patients.

Ambulatory Care↗

Chemical and biological pollution contribute to the immunological profiles of free-ranging harbor seals.

Polychlorinated biphenyls and other persistent organic pollutants have been associated with immunotoxicity and outbreaks of (infectious) disease in marine mammals by rendering them vulnerable to infection by pathogens such as viruses and bacteria. In an immunotoxicological study of free-ranging harbor seals (Phoca vitulina), we obtained samples of blood and blubber from seal pups that were live-captured from two remote and two near-urban sites in British Columbia, Canada, and Washington state, USA. Using these samples, we quantified hematology, innate immune function, adaptive immune function, and polychlorinated biphenyl accumulation. While controlling for confounding factors (age, sex, and condition), univariate correlations between phagocytosis (r2 = 0.30, p = 0.002), respiratory burst (r2 =0.45, p= 0.000), T-lymphocyte function (r2 = 0.16, p = 0.028), lymphocyte signaling (r2 = 0.17, p = 0.025), and lymphocyte counts (r2 = 0.29, p = 0.002), and polychlorinated biphenyl concentrations suggested chemical-associated immunotoxicity. Principal component analysis of immunological endpoints provided additional evidence of immunotoxic effects in seals. However, principal component analysis also identified a noncontaminant-related factor by distinguishing between seals inhabiting urban versus remote sites, with results being consistent with increased pathogen exposure. Elevated fecal coliform concentrations in water, and observations of terrestrial spill-over pathogens in local seals, further support the notion of biological pollution at these sites. Although our study highlights the role that environmental contaminants might play in rendering marine mammal populations vulnerable to disease through immunotoxicity, it also suggests that biological pollution represents an emerging conservation concern.

Animals↗

Measurement of mouse urinary type IV collagen using time-resolved fluoroimmunoassay.

Urinary level of type IV collagen is an important indicator for early renal dysfunction, but there has been no practical system to measure mouse type IV collagen adaptable to extremely small amounts of urine samples. We developed a highly sensitive time-resolved fluoroimmunoassay (TR-FIA) to measure mouse urinary type IV collagen. Based on the structural features of type IV collagen molecule, dithiothreitol (DTT) was used for pretreatment of the samples. This assay permits measurement of 100 pg/ml type IV collagen in 5 microl urine samples. Urinary levels of type IV collagen derived from 12 samples of two different mouse strains (KK/Ta and BALB/c) were measured using this assay. The results demonstrated very clearly the difference in values of urinary type IV collagen between diabetic mice and non-diabetic mice. Compared with the conventional enzyme-linked immunosorbent assay (ELISA), this method requires far smaller volumes of samples, and is best suited to mouse models in future experiments.

Animals↗

Reliability and factor structure of the Brazilian version of the Center for Epidemiologic Studies-Depression.

The Center for Epidemiologic Studies-Depression scale is a screening test designed by the National Institute of Mental Health (USA) to screen for 'depressed' individuals in the community. It has been widely used in different settings and cultures displaying variable psychometric properties. The present article describes coefficients of internal consistency and factor analysis of the CES-Depression version adapted for use in Brazil. In a sample of 523 college students internal consistency measured by Cronbach alpha was .85. Structure analysis evidenced that two questions were poorly correlated with the remaining questions. Factor analysis resulted in a four-dimension solution accounting for 53.8% of the total item variance. The scale is adequate for use in this different cultural setting.

Adolescent↗

Influence of the amount of dietary fiber on the available energy from hindgut fermentation in growing pigs: use of cannulated pigs and in vitro fermentation.

Available energy from hindgut fermentation to pigs fed various amounts of dietary fiber was investigated using an in vivo-in vitro methodology. Six growing pigs fitted with a simple T-shaped cannula at the terminal ileum, and following a Latin-square design, were fed 3 diets differing in the content of non-starch polysaccharides (NSP): a low fiber diet (LFD, 77 g/kg of DM), a standard fiber diet (SFD, 160 g/kg of DM), and a high fiber diet (HFD, 240 g/kg of DM). After adaptation to the diet for 10 d, samples from feces and ileum were collected and analyzed for DM, energy, NSP, and chromic oxide; feces were also analyzed for short chain fatty acids (SCFA). Freeze-dried ileal samples (10 g/L) were fermented in vitro in a fecal slurry consisting of an anaerobic mineral salt medium and feces (50 g/L) from cannulated pigs fed the same diets. Available energy was calculated from the amount of SCFA produced in vitro after 48 h of incubation. Nonstarch polysaccharide content in the fermented material was measured to assess the in vitro degradation of this fraction. Increasing dietary NSP from 77 to 240 g/kg of feed DM increased (P < 0.001) ileal flow from 199 to 468 g/kg of feed, leading to a reduction in the energy digested at the terminal ileum, from 15 to 11 MJ/kg of feed DM and an increment in energy digested in the hindgut, from 1.6 to 3.5 MJ/kg of feed DM. Total in vitro production of SCFA/kg of feed DM was dependent on the amount of ileal substrate available for fermentation; that is, increased concentrations of NSP in the diet led to an increase in the SCFA that may be available to the animal (P < 0.001). The molar ratio of SCFA produced in vitro was affected by diet; the high fiber diet showed the greatest (P = 0.004) proportion of acetic acid, and the low fiber diet showed a tendency (P = 0.081) to an increased butyric acid proportion compared with the other 2 diets. Net disappearance of NSP during fermentation in vivo and in vitro were compared and showed a close relationship (P < 0.001, slope = 0.906, r = 0.960). In our experimental conditions, available energy as SCFA to the animal from hindgut fermentation increased with the concentration of dietary NSP (P < 0.001) and provided between 7.1 and 17.6% of the total available energy.

Animal Feed↗

Collagen deficiency and ruptured cerebral aneurysms. A clinical and biochemical study.

Skin and temporal arterial biopsies were obtained from 17 patients undergoing surgery for ruptured cerebral aneurysm, and specimens were taken from six age- and sex-matched control surgical patients. Radioactively labeled and control tissue collagen patterns were studied by interrupted polyacrylamide gel electrophoresis (PAGE), using the trisborate buffer system or by carboxymethyl cellulose (CMC) chromatography. Type III/I collagen ratios were then measured from autoradiographs of the radioactively labeled samples using the Joyce Loebl gel scanner adapted for flat bed gels. In the case of the CMC labeled material, the ratios were measured by the ratios of the summed radioactively labeled alpha 1(III), alpha 2(II), and alpha 2(I) peaks. Eleven of the 17 patients were Type III collagen-deficient while all of the six control patients had normal collagen ratios. The implications of these findings are discussed.

Adult↗

Field studies of a rapid, accurate means of quantifying Schistosoma haematobium eggs in urine samples.

A new means of quantifying schistosome eggs has been adapted from the laboratory to the field. Urine from 510 individuals in areas of Kenya with a high prevalence of schistosomiasis haematobia was injected in aliquots of 10, 5 and 1 ml on to transparent Nuclepore filters, 13 mm in diameter. The filters were placed face down on glass slides and were read without staining at x40 magnification. The method has been shown to be accurate, sensitive, and reproducible, and also extremely rapid.

Adolescent↗

[Validation of the French version of the Raine Schizotypal Personality Disorder Questionnaire--categorial and dimensional approach to schizotypal personality traits in a normal student population].

Most of existing self-report measures of schizotypal personality assess only few of the nine traits of Schizotypal Personality Disorder (SPD) according to DSM III-R or DSM IV. The Raine's Schizotypal Personality Questionnaire (SPQ) is one of the most widely used questionnaire for SPD diagnosis. This well adapted tool for screening SPD in large samples from general population allows simple and quick evaluations by the mean of 74 items and nine sub-scales exploring DSM IV criteria of the trouble. With the original sample of American students, Raine found SPQ to have high internal reliability (0.91) and reported cutoffs for the top and the bottom ten percents of SPQ scores at respectively 41/74 and 12/74. We reported here the validation study of the French version of the SPQ in a sample of 232 students (males: n = 107; females: n = 125; mean age: 21.17 +/- 1.47; mean educational level: 13.36 +/- 1.06). The French version has high internal reliability (SPQ total: Cronbach's alpha = 0.91; SPQ nine subscales: Cronbach's alpha = 0.57 to 0.76). The ten percent high and low cutoff scores on the distribution of SPQ scores are respectively 40/74 and 7/74 for the total sample. However, gender differences are observed: 38/74 and 9/74 for females; 42/74 and 6/74 for males. A Principal Component Analysis (PCA) confirms the high internal reliability. Moreover, PCA evidences a three-factor model of schizotypy reflecting "positive or cognitive-perceptual", "negative or social-interpersonal", and "disorganization" latent factors. These results replicate previous works on the topic with the same instrument.

Adolescent↗

Epidemiological and clinical study of autistic children in Yogyakarta, Indonesia.

This epidemiological study of autistic children was carried out in Yogyakarta, Indonesia. The approaches considered included the analysis of cross-sectional data on non-verbal intelligence, social maturity behavior or adaptive skills test by using statistical or sampling controls. The 5,120 children studied were all Javanese. Doctors, nurses of Public Health Centers and school teachers assisted in assessing the above subjects. The Bryson's screening scale for evaluating autistic children was used. A total of 66 children who scored above 16 were identified. Final diagnosis was done by means of CARS. Six children, who lived within Yogyakarta Municipality, were identified. The prevalence rate of autism within the birth cohort who were born between June 1984 and May 1991 was 12 per 10,000. Observations of the social interactions of 6 autistic and 6 normal children of equivalent mental age were carried out during a free play situation and reunion situation with mother and a stranger. A comparison of the two groups during social interaction made it clear that the autistic children showed more attachment, directed more social-interaction and more physical contact toward their mothers than to strangers during the reunion situation.

Autistic Disorder↗

[Some aspects of intraspecific variability of the closely related species of the Dermacentor marginatus complex (Acari: Ixodidae) as demonstration of microevolutionary process].

Individual and geographical variability of male conscutum pattern in five closely related species of the Dermacentor marginatus complex (D. marginatus, D. ushakovae, D. niveus, D. silvarum, D. nuttalli) has been examined. The pattern has been examined in three to six geographical samples within each specific distribution range. It was established that the pattern of the conscutum central area is subjected to individual and geographical variability on an extreme degree and gives very few intraspecific differential characters, but it shows interspecific differences in some cases. However the dark patches of marginal flank (npkappadelta and 3deltakappa respectively, see Fig. 3) show statistically significant differences in length in all geographical samples of the first four species. Characteristic features of ixodid ticks morpogenesis rule out to a considerable extent the possibility to trace geographical variability of the same characters by all stages of ontogenesis. In these cases, when it is possible, the tendencies to variability on different stages do not always coincide. There is no coordination within the species in the degree of differences of examined geographical samples by pattern and morphological characters, i. e. very complex polymorphis is observed. In the context of the evolution that kind of polymorphism of the geographical samples has provided the species with high adaptive potential for conquest of the extensive range and maintaining large numbers. The question of allopatric speciation of the closely related D. marginatus-D. silvarum with small secondary area of transgression their ranges, and sympatric speciation of the D. ushakovae-D. niveus is postulated.

Animals↗

Quantitative analysis of platelet activating factor treated with pentafluorobenzoyl chloride using gas chromatography/negative ion chemical ionization mass spectrometry.

To confirm that platelet activating factor (PAF) plays an important role as a mediator in acute inflammation and allergic reaction, it is necessary to develop a more sensitive and stable method for measuring trace amounts of PAF in various biological samples. For this reason, the authors have adapted gas chromatography/negative ion chemical ionization mass spectrometry (GC/NICI-MS) (developed by Ramesha and Pickett in 1985) by employing an SPB-1 column and isobutane as the reagent gas. Furthermore, with this method the authors attempted an investigation of the time course of hexadecyl- and octadecyl-PAF production and release from human polymorphonuclear leukocytes (PMNs) stimulated by Ca-ionophore A23187. PMNs were obtained from venous blood from a human donor by centrifugation and were stimulated by 5 microM of Ca-ionophore A23187. PAF was purified using a SEP-PAK silica column and thin layer chromatography, and then was hydrolyzed with phospholipase C. The extract was treated with pentafluorobenzoyl chloride. Using 1-O-hexadecyl-2-acetyl (perdeuterated)-sn-phosphocholine) as an internal standard, the following results were obtained: The standard curve for this quantitative analysis was linear with a correlation coefficient of 0.9997 from 1 pg to 200 ng. The authors assumed that quantities as low as a few picograms of PAF could be measured by the method. The production of hexadecyl-PAF in the cell pellet peaked (8.1 +/- 1.34 ng/10(7) cells) at 2 min after stimulation and that in the supernatant peaked (6.3 +/- 0.97 ng/10(7) cells) at approximately 7 min after stimulation. Octadecyl-PAF could not be detected in this experiment.(ABSTRACT TRUNCATED AT 250 WORDS)

Benzoates↗