Estimating motor unit numbers from a CMAP scan: Repeatability study on three muscles at 15 centres.

OBJECTIVE: To assess the repeatability and suitability for multicentre studies of MScanFit motor unit number estimation (MUNE), which involves modelling compound muscle action potential (CMAP) scans. METHODS: Fifteen groups in 9 countries recorded CMAP scans twice, 1-2 weeks apart in healthy subjects from abductor pollicis brevis (APB), abductor digiti minimi (ADM) and tibialis anterior (TA) muscles. The original MScanFit program (MScanFit-1) was compared with a revised version (MScanFit-2), designed to accommodate different muscles and recording conditions by setting the minimal motor unit size as a function of maximum CMAP. RESULTS: Complete sets of 6 recordings were obtained from 148 subjects. CMAP amplitudes differed significantly between centres for all muscles, and the same was true for MScanFit-1 MUNE. With MScanFit-2, MUNE differed less between centres but remained significantly different for APB. Coefficients of variation between repeats were 18.0% for ADM, 16.8% for APB, and 12.1% for TA. CONCLUSIONS: It is recommended for multicentre studies to use MScanFit-2 for analysis. TA provided the least variable MUNE values between subjects and the most repeatable within subjects. SIGNIFICANCE: MScanFit was primarily devised to model the discontinuities in CMAP scans in patients and is less suitable for healthy subjects with smooth scans.

Effects of buprenorphine on acute pain and inflammation in the adjuvant-induced monoarthritis rat model.

Background and aim: Animal modelling of arthritis is often associated with pain and suffering. Severity may be reduced with the use of analgesia which is, however, often withheld due to concerns of introducing a confounding variable. It is therefore important to design and validate pain relief protocols that reduce pain without compromising the scientific objectives. The present study evaluated the effect of buprenorphine analgesia in the immediate post-induction period of an adjuvant-induced monoarthritic rat model. The aim of this study was to extend previous work on refinement of the model by alleviating unnecessary pain. Methods: Male and female Sprague Dawley rats were injected with 20 µl of complete Freund's adjuvant (CFA) into the left ankle. Rats were treated with buprenorphine, either injected subcutaneously or ingested voluntarily, and were compared to rats given subcutaneous injections with vehicle (saline or pure nut paste) or carprofen the first three days post CFA-injection. Measurements of welfare, clinical model-specific parameters and pain-related behaviour were assessed. Results: Buprenorphine, administered either subcutaneously (0.10 or 0.15 mg/kg, twice daily) or by voluntary ingestion in nut paste (1.0 or 3.0 mg/kg, twice daily), improved mobility, stance, rearing and lameness scores significantly 7 h post CFA-injection. Mechanical hyperalgesia peaked at 7 h and was significantly lower in buprenorphine-treated animals, compared to vehicle-treated animals. Joint circumference was highest 24-72 h after CFA injection. Animals treated with buprenorphine did not decrease in joint circumference, opposite carprofen treated animals. Conclusion: Buprenorphine, administered either subcutaneously or by voluntary ingestion, provides adequate analgesia for both sexes within the first 24 h post CFA-injection. Buprenorphine treatment improved clinical scores and appeared not to suppress the inflammatory response. The present study supports previous findings that voluntarily ingested buprenorphine is an effective alternative to repeated injections.

[Prevalence of asthma and associated risk factors in schoolchildren from an area of indigenous population]. / Prevalencia de asma y factores de riesgo asociados en escolares provenientes de una zona de pueblos indígenas.

INTRODUCTION: The prevalence of asthma in Chile ranges from 10.2 to 14.9%. In previous studies, the Araucanía Region has not been included. OBJECTIVE: To determine the prevalence of asthma in the school po pulation of the Araucanía Region. SUBJECTS AND METHOD: The ISAAC questionnaires were used for asthma symptoms in addition to a questionnaire for recording sociodemographic data, belonging to the Mapuche ethnicity, type of heating, exposure to indoor air pollution, and family history. The Binomial Regression Model was used to evaluate the effect of each of the different variables of inter est, adjusting by age groups (6-7 and 13-14 years). The model also evaluated the additive interaction between these variables and age. RESULTS: 823 surveys were applied, where the prevalence of asthma was 14.2% and 23.2% in the 6-7 year old group and the 13-14 year old group, respectively. 43.7% de clare to be Mapuche, 32.9% live in rural areas, and 81.4% use firewood as a heating method. Through binomial regression model analysis, where for the study group, living in the coastal commune was significantly associated with a lower prevalence of asthma (-13.2% [CI: -17.7 to -8.6]. The history of an asthmatic mother was significantly associated with a higher prevalence of asthma (17.9% [CI: 2.7 to 33.1]. Living in a rural area, to be Mapuche or the type of heating used, showed differences statistically significant (p 0.18, p 0.609 and p 0.480, respectively) Conclusion: 13-14 year-olds school children in the Araucanía Region presented a higher prevalence of asthma. There was an association with asthma in the mother where this increase is not associated with living in a rural area, to be Ma puche or type of heating.

Liver Transplant After Normothermic Regional Perfusion From Controlled Donors After Circulatory Death: The Norwegian Experience.

BACKGROUND: In order to meet the increasing demand for donor organs, the concept of donation after circulatory death (DCD) was reintroduced in Norway, first as a pilot study, followed by the use of DCD as institutional practice. We report the current Norwegian experience with liver transplant after DCD. METHODS: After acceptance from next of kin, life support was withdrawn from patients with devastating brain injury and cardiac arrest observed. After a 5-minute "no-touch" period, extracorporeal membrane oxygenation for post mortem normothermic regional perfusion (NRP) by extracorporeal membrane oxygenator circuit was established. Data from all liver transplant recipients receiving controlled DCD (cDCD) livers in Oslo were analyzed. RESULTS: From 2015 to 2017, a total of 8 patients underwent liver transplant with cDCD and NRP liver grafts in Norway. Median Model for End-Stage Liver Disease score was 26 (range, 6-40). There were no cases of delayed graft function or graft loss. Seven patients have reached 1 year of follow-up, and 1 patient has reached 6 months. Two patients have recurrence of primary disease (primary sclerosing cholangitis and steatohepatitis). All patients had normalized liver function at last follow-up. Two patients underwent procedures for biliary complications. In 1 patient, leakage from the cystic duct was successfully handled endoscopically by stenting. In the other patient, a suspected stricture on magnetic resonance imaging led to an endoscopic retrograde cholangiopancreatography, which did not confirm signs of biliary stenosis. There was 1 instance of hepatic artery stenosis, which was managed with endovascular technique. CONCLUSION: The results after liver transplant using cDCD with NRP are good. The rate of complications seems to be within the same range as when using conventional donation after brain death grafts.

Insulin resistance in vascular endothelial cells promotes intestinal tumour formation.

The risk of several cancers, including colorectal cancer, is increased in patients with obesity and type 2 diabetes, conditions characterised by hyperinsulinaemia and insulin resistance. Because hyperinsulinaemia itself is an independent risk factor for cancer development, we examined tissue-specific insulin action in intestinal tumour formation. In vitro, insulin increased proliferation of intestinal tumour epithelial cells by almost two-fold in primary culture of tumour cells from ApcMin/+ mice. Surprisingly, targeted deletion of insulin receptors in intestinal epithelial cells in ApcMin/+ mice did not change intestinal tumour number or size distribution on either a low or high-fat diet. We therefore asked whether cells in the tumour stroma might explain the association between tumour formation and insulin resistance. To this end, we generated ApcMin/+ mice with loss of insulin receptors in vascular endothelial cells. Strikingly, these mice had 42% more intestinal tumours than controls, no change in tumour angiogenesis, but increased expression of vascular cell adhesion molecule-1 (VCAM-1) in primary culture of tumour endothelial cells. Insulin decreased VCAM-1 expression and leukocyte adhesion in quiescent tumour endothelial cells with intact insulin receptors and partly prevented increases in VCAM-1 and leukocyte adhesion after treatment with tumour necrosis factor-α. Knockout of insulin receptors in endothelial cells also increased leukocyte adhesion in mesenteric venules and increased the frequency of neutrophils in tumours. We conclude that although insulin is mitogenic for intestinal tumour cells in vitro, impaired insulin action in the tumour microenvironment may be more important in conditions where hyperinsulinaemia is secondary to insulin resistance. Insulin resistance in tumour endothelial cells produces an activated, proinflammatory state that promotes tumorigenesis. Improvement of endothelial dysfunction may reduce colorectal cancer risk in patients with obesity and type 2 diabetes.

High environmental temperature around farrowing induced heat stress in crated sows.

The aim of the experiment was to study the impact of high ambient temperature (25°C) around farrowing on crated sows unable to perform thermoregulatory behavior. Twenty sows were housed in 2 farrowing rooms in conventional farrowing crates. In 1 room (CONTROL) temperature was kept at 20°C. In the other room (HEAT) temperature was initially kept at 20°C and gradually raised until it reached 25°C from d 112 to 115 of gestation. Then the temperature was gradually lowered to 20°C. Sows were continuously video recorded for behavior recording. Sows' respiration rates were recorded from d 3 before farrowing to d 5 after farrowing. Sows' rectal temperatures were recorded from d 1 before farrowing to d 8 after farrowing, and sows' udder surface temperatures were recorded from the day of farrowing to d 3 after farrowing. All measures were recorded daily. Sows' BW were recorded at d 108 of gestation and at weaning. Sows' back fat was recorded on farrowing day, when room temperature was set again at 20°C, and at weaning. Piglets were weighed at d 1, 14, and 21. The HEAT sows spent a higher proportion of time lying in the lateral position than CONTROL sows, both during the 16 h before farrowing and the 24 h after the start of farrowing ( < 0.05), but with no difference in the amount of time spent lying down between groups ( > 0.10). The HEAT sows had higher rectal temperature on d 1 after farrowing ( < 0.05) and had udder surface temperature 0.9°C higher than that of CONTROL sows during the recording period ( < 0.05). The HEAT sows also tended to have longer farrowing duration ( < 0.10). Respiration rate was higher in HEAT sows on d 1 before farrowing and on the day of farrowing. On d 7, 8, and 9, CONTROL sows had higher feed intake ( < 0.05), and piglets from CONTROL sows were heavier at d 21 after farrowing ( < 0.05). High ambient temperature around farrowing altered sows' postural behavior. Sows reacted to the thermal challenge with higher respiration rate around farrowing, but both their rectal and udder temperatures were elevated, indicating that they were not able to compensate for the higher ambient temperature. High ambient temperature negatively influenced sows' feed intake, with negative impact on piglets' weaning weight. High temperatures around farrowing (25°C) compromise crated sows' welfare, with a potential negative impact on offspring performance.

Transcriptome Analysis of Aspergillus flavus Reveals veA-Dependent Regulation of Secondary Metabolite Gene Clusters, Including the Novel Aflavarin Cluster.

The global regulatory veA gene governs development and secondary metabolism in numerous fungal species, including Aspergillus flavus. This is especially relevant since A. flavus infects crops of agricultural importance worldwide, contaminating them with potent mycotoxins. The most well-known are aflatoxins, which are cytotoxic and carcinogenic polyketide compounds. The production of aflatoxins and the expression of genes implicated in the production of these mycotoxins are veA dependent. The genes responsible for the synthesis of aflatoxins are clustered, a signature common for genes involved in fungal secondary metabolism. Studies of the A. flavus genome revealed many gene clusters possibly connected to the synthesis of secondary metabolites. Many of these metabolites are still unknown, or the association between a known metabolite and a particular gene cluster has not yet been established. In the present transcriptome study, we show that veA is necessary for the expression of a large number of genes. Twenty-eight out of the predicted 56 secondary metabolite gene clusters include at least one gene that is differentially expressed depending on presence or absence of veA. One of the clusters under the influence of veA is cluster 39. The absence of veA results in a downregulation of the five genes found within this cluster. Interestingly, our results indicate that the cluster is expressed mainly in sclerotia. Chemical analysis of sclerotial extracts revealed that cluster 39 is responsible for the production of aflavarin.

A strong response to selection on mass-independent maximal metabolic rate without a correlated response in basal metabolic rate.

Metabolic rates are correlated with many aspects of ecology, but how selection on different aspects of metabolic rates affects their mutual evolution is poorly understood. Using laboratory mice, we artificially selected for high maximal mass-independent metabolic rate (MMR) without direct selection on mass-independent basal metabolic rate (BMR). Then we tested for responses to selection in MMR and correlated responses to selection in BMR. In other lines, we antagonistically selected for mice with a combination of high mass-independent MMR and low mass-independent BMR. All selection protocols and data analyses included body mass as a covariate, so effects of selection on the metabolic rates are mass adjusted (that is, independent of effects of body mass). The selection lasted eight generations. Compared with controls, MMR was significantly higher (11.2%) in lines selected for increased MMR, and BMR was slightly, but not significantly, higher (2.5%). Compared with controls, MMR was significantly higher (5.3%) in antagonistically selected lines, and BMR was slightly, but not significantly, lower (4.2%). Analysis of breeding values revealed no positive genetic trend for elevated BMR in high-MMR lines. A weak positive genetic correlation was detected between MMR and BMR. That weak positive genetic correlation supports the aerobic capacity model for the evolution of endothermy in the sense that it fails to falsify a key model assumption. Overall, the results suggest that at least in these mice there is significant capacity for independent evolution of metabolic traits. Whether that is true in the ancestral animals that evolved endothermy remains an important but unanswered question.

Caveolin expression changes in the neurovascular unit after juvenile traumatic brain injury: signs of blood-brain barrier healing?

Traumatic brain injury (TBI) is one of the major causes of death and disability in pediatrics, and results in a complex cascade of events including the disruption of the blood-brain barrier (BBB). A controlled-cortical impact on post-natal 17-day-old rats induced BBB disruption by IgG extravasation from 1 to 3 days after injury and returned to normal at day 7. In parallel, we characterized the expression of three caveolin isoforms, caveolin 1 (cav-1), caveolin 2 (cav-2) and caveolin 3 (cav-3). While cav-1 and cav-2 are expressed on endothelial cells, both cav-1 and cav-3 were found to be present on reactive astrocytes, in vivo and in vitro. Following TBI, cav-1 expression was increased in blood vessels at 1 and 7 days in the perilesional cortex. An increase of vascular cav-2 expression was observed 7 days after TBI. In contrast, astrocytic cav-3 expression decreased 3 and 7 days after TBI. Activation of endothelial nitric oxide synthase (eNOS) (via its phosphorylation) was detected 1 day after TBI and phospho-eNOS was detected both in association with blood vessels and with astrocytes. The molecular changes involving caveolins occurring in endothelial cells following juvenile-TBI might participate, independently of eNOS activation, to a mechanism of BBB repair while, they might subserve other undefined roles in astrocytes.

Investigating the long-term effect of subchronic phencyclidine-treatment on novel object recognition and the association between the gut microbiota and behavior in the animal model of schizophrenia.

Subchronic phencyclidine (subPCP) treatment induces schizophrenic-like behavior in rodents, including cognitive deficits and increased locomotor sensitivity towards acute administration of PCP. Evidence is accumulating that the gut microbiota (GM) influences behavior through modulation of the microbiota-gut-brain axis, and hence, part of the variation within this animal model may derive from variation in the GM. The aims of this study was to investigate first, the duration of subPCP-induced cognitive impairment in the novel object recognition test, and second, the possible effect of subchronic PCP-treatment on the GM, and the association between the GM and the behavioral parameters. The association was further investigated by antibiotic reduction of the GM. Male Lister Hooded rats were dosed twice daily i.p. with either 5mg/kg PCP or sterile isotonic saline for seven days followed by a seven-day washout period. Rats were tested in the novel object recognition and the locomotor activity assays immediately after, three weeks after, or six weeks after washout, and the fecal GM was analyzed by high throughput sequencing. Antibiotic- and control-treated rats were tested in the same manner following washout. In conclusion, subPCP-treatment impaired novel object recognition up to three weeks after washout, whereas locomotor sensitivity was increased for at least six weeks after washout. Differences in the core gut microbiome immediately after washout suggested subPCP treatment to alter the GM. GM profiles correlated to memory performance. Administration of ampicillin abolished the subPCP-induced memory deficit. It thus seems reasonable to speculate that the GM influences memory performance, contributing to variation within the model.

Fasting of mice: a review.

Fasting of mice is a common procedure performed in association with many different types of experiments mainly in order to reduce variability in investigatory parameters or to facilitate surgical procedures. However, the effects of fasting not directly related to the investigatory parameters are often ignored. The aim of this review is to present and summarize knowledge about the effects of fasting of mice to facilitate optimization of the fasting procedure for any given study and thereby maximize the scientific outcome and minimize the discomfort for the mice and hence ensure high animal welfare. The results are presented from a number of experimental studies, providing evidence for fasting-induced changes in hormone balance, body weight, metabolism, hepatic enzymes, cardiovascular parameters, body temperature and toxicological responses. A description of relevant normal behaviour and standard physiological parameters is given, concluding that mice are primarily nocturnal and consume two-thirds of their total food intake during the night. It is argued that overnight fasting of mice is not comparable with overnight fasting of humans because the mouse has a nocturnal circadian rhythm and a higher metabolic rate. It is suggested that because many physiological parameters are regulated by circadian rhythms, fasting initiated at different points in the circadian rhythm has different impacts and produces different results.

The prevalence of couple infertility in the United States from a male perspective: evidence from a nationally representative sample.

Infertility is a couple-based fecundity impairment, although population level research is largely based upon information reported by female partners. Of the few studies focusing on male partners, most focus on the utilization of infertility services rather than efforts to estimate the prevalence and determinants of infertility as reported by male partners. Data from a nationally representative sample of men aged 15-44 years who participated in the 2002 National Survey of Family Growth (NSFG) were used to estimate the prevalence of infertility and determinants of longer time-to-pregnancy (TTP) using the novel current duration (CD) approach. Using backward recurrence time parametric survival methods, we estimated infertility prevalence (TTP > 12 months) and time ratios (TR) associated with TTP as derived from males' reported CD of their pregnancy attempt. The estimated prevalence of infertility was 12.0% (95% CI: 7.0, 23.2). Longer TTP was associated with older male age (35-45 vs. 17-24 years) (TR: 2.49; 95% CI: 1.03, 6.03), biological childlessness (TR: 1.53; 95% CI: 1.07, 2.19) and lack of health insurance (TR: 1.73; 95% CI: 1.02, 2.94) after controlling for the differences in couples' age and other socioeconomic factors. The prevalence of infertility based on male reporting is consistent with estimates of infertility in the US found in prospective cohort studies and CD studies based on female reporting. Our findings suggest that male partners can reliably inform about couple infertility. Interventions and services aimed at reducing couple infertility should include attention to male factors associated with longer TTP identified in this study.

Cumulative incidence rate of medical consultation for fecundity problems--analysis of a prevalent cohort using competing risks.

STUDY QUESTION: What is the incidence of medical consultation for fecundity problems in the French population, taking into account pregnancy occurrence and resumption of contraceptive use?. SUMMARY ANSWER: Considering the occurrence of a pregnancy and resumption of use of contraception as competing risks, the cumulative incidence rate of medical consultation for fecundity problems was 9.0% [95% confidence interval (CI): 6.5%; 11.9%] after 12 months of unprotected intercourse and 12.2% [95% CI: 9.6%; 15.3%] after 24 months. WHAT IS KNOWN ALREADY: Estimates of the prevalence of medical consultation due to involuntary infertility among couples who have sought a pregnancy for more than 12 months range from 25 to 50%. Most of the studies however are limited by retrospective data collection, without considering the duration of time since the beginning of the period of unprotected intercourse (PUI) and without considering medical consultation for fecundity problems as a competing risk. STUDY DESIGN, SIZE, DURATION: This study is based on the Observatory of Fecundity in France survey, a population-based probability survey designed to estimate the frequency of involuntary infertility on a nationwide basis and to explore the associations with environmental factors. Women answered two telephone questionnaires, the first at the time of enrolment in 2007, the second at follow-up 1 year later. The current analysis was performed among a subsample of 6577 women recruited before or during a PUI and followed-up for 1 year. PARTICIPANTS/MATERIALS, SETTING, METHODS: The study sample comprised 940 women aged 18-44 years who had a PUI between the time of enrolment and the 1-year follow-up, and who had not consulted a physician for fecundity problems for the current PUI prior to enrolment. Women reported all the medical consultations they had because of difficulties becoming pregnant during the current PUI. The date of each consultation was carefully assessed. In France, women can consult a gynaecologist directly without referral by their general practitioner. The occurrence of a pregnancy and resumption of contraceptive use were considered as informative censoring events, using a competing risk model. MAIN RESULTS AND THE ROLE OF CHANCE: Using the competing risk survival model, the cumulative incidence rate of first consultation was 9.0% [95% CI: 6.5%; 11.9%] 12 months after the start of the PUI and 12.2% [95% CI: 9.6%; 15.3%] after 24 months. The Kaplan-Meier method, which does not take competing risks into account, yielded substantially higher estimates: 26.0% [95% CI: 18.8%; 32.5%] at 12 months and 56.8% [95% CI: 44.2%; 66.6%] at 24 months. Among the 219 women who had attempted to become pregnant for at least 12 months, cumulative incidences of first medical consultations were 28.2% [18.7-38.9%] 24 months after the start of the PUI, and 31.2% [21.3- 42.4%] after 36 months. The rates were higher among nulliparous but non-nulligravid women, followed by nulligravid women, as compared with parous women. Age was not strongly related to the occurrence of medical consultation. LIMITATIONS, REASONS FOR CAUTION: The main limitation of this study is the number of women lost to follow-up (29.7%). In addition, results regarding the absence of an age effect should be taken with caution as few women in our study were aged over 35 years. Although such an attrition rate is commonly observed in prospective studies in the general population, it could have induced a selection bias that may have led to an underestimation of the rates of medical consultation. Sensitivity analyses, using the inverse probability weighting method suggest that our results are unlikely to be biased. WIDER IMPLICATIONS OF THE FINDINGS: This study reveals frequencies of medical consultation for fertility problems, which, after considering competing events such as pregnancy in a relevant statistical model, are lower than generally reported in the literature. The results also indicate the existence of a difference between the potential need and the actual use of medical care for fecundity problems. This suggests a need for studies to look for factors other than medical recommendations that may play a role in the patterns of medical seeking behaviours for fecundity problems, such as women's reproductive history, socio-economic characteristics or accessibility to infertility services. STUDY FUNDING/COMPETING INTEREST(S): The study was funded by grants from ANR (French Agency for Research, SEST call on Environmental and Occupational Health), ANSES (French Agency for Food, environmental and Occupational Health Safety, EST call on Environmental and Occupational Health), InVS (French Institute for Public Health Surveillance). The team of Environmental Epidemiology applied to Fecundity and Reproduction has been funded by an AVENIR grant from Inserm (2007). Authors declare no conflict of interest.

Estimation of the frequency of involuntary infertility on a nation-wide basis.

BACKGROUND: Assessing couple fecundity on a nation-wide basis without excluding couples who eventually remain infertile is challenging. Our aim was to describe couple fecundity (in terms of frequency of involuntary infertility) among the general population living in France. METHODS: We used a current-duration design. A random sample of 64 262 households was selected in 2007-2008, allowing us to identify 15 810 women aged 18-44 years. Eligible women (n= 1089) were those having regular sexual intercourse with a male partner, not using any method of contraception and not having delivered in the previous 3 months. These women reported information on the current duration of unprotected intercourse (CDUI, the time elapsed between the start of the period of unprotected intercourse and the time of inclusion in the study). The CDUI distribution was used to estimate the frequency of involuntary infertility, using a newly developed statistical technique that does not require couples to be followed up until the end of the period of unprotected intercourse. RESULTS: CDUI was defined for 867 women. An estimated 46% of couples had no detected pregnancy conceived during the first 6 months of unprotected intercourse [95% confidence interval (CI), 36-56%]. The proportions of couples with no detected pregnancy within 12 and 24 months were 24% (19-30%) and 11% (8-14%), respectively. CONCLUSIONS: These results constitute one of the few descriptions of the fecundity of a nation-wide representative sample of couples from the general population, not limited to couples who eventually conceived or to those resorting to medical help.

Less invasive blood sampling in the animal laboratory: clinical chemistry and haematology of blood obtained by the Triatominae bug Dipetalogaster maximus.

Dipetalogaster maximus (Dipmax), a blood-sucking bug belonging to the family Reduviidae, has been used to obtain blood samples, for example for clinical chemistry and haematology, in a variety of zoo animals and wildlife. Using this bug allows stress-free blood sampling as the bug is able to draw blood without the mammal noticing the bug. In laboratory animal science, the need for blood samples from unstressed animals may arise, especially in animal behaviour research. The use of Dipmax bugs may prove a valuable tool for this purpose. To validate the method, we compared an array of standard blood parameters sampled from New Zealand White rabbits, sampled either by the use of bugs or by the conventional method; puncture of vena auricularis caudalis. The overall hypothesis was that there was no significant difference in clinical chemistry and haematological parameters between the bug method and the conventional method. A total of 17 clinical parameters as well as 12 haematological parameters were measured and compared in New Zealand White rabbits. The results showed that for 13 of these 29 analysed parameters, the bug method and the conventional method did not give significantly different results, and the obtained results were thus directly comparable. For the remaining parameters the obtained results were significantly different. However, all parameters were measurable in the bug samples. The influences of the bug metabolism on these parameters are discussed.

Pathology, tissue metalloproteinase transcription and haptoglobin responses in mice after experimental challenge with different isolates of Pasteurella multocida obtained from cases of porcine pneumonia.

Pasteurella multocida is a major cause of porcine pneumonia, but the pathogenesis of the disease is poorly defined. The aim of this study was to further understand the host response to infection by use of a mouse model of P. multocida pneumonia. Twenty female mice were divided into four groups (n=5). Three groups were infected with one of three isolates of P. multocida isolated from clinical cases of chronic porcine pneumonia with necrotizing, suppurative and non-suppurative lesions, respectively. The fourth group served as uninfected controls. Mice were killed 24 h postinfection and samples were collected for bacteriology, histopathology and in-situ hybridization for detection of P. multocida. Measurements of expression of genes encoding matrix metalloproteinase 9 (MMP9) and tissue inhibitor of metalloproteinase 1 (TIMP1) in lung tissue and quantification of serum haptoglobin concentration were performed. P. multocida was found in the lung and spleen. Lung lesions were characterized by deposition of fibrin in alveoli and bronchioles, perivascular oedema, suppuration and necrosis. The cellular infiltration was mainly of neutrophils. Splenic neutrophilic infiltration was also evident. Minor differences in the severity and nature of lesions were seen according to the isolate of P. multocida used for infection. Intranasal infection of mice can therefore be used to evaluate the host response and lesions caused by P. multocida obtained from porcine pneumonic infections. The inflammatory response in this model is associated with increased tissue expression of genes encoding MMP9, TIMP1 and serum haptoglobin concentration.

GSEVM v.2: MCMC software to analyze genetically structured environmental variance models.

This note provides a description of software that allows to fit Bayesian genetically structured variance models using Markov chain Monte Carlo (MCMC). The gsevm v.2 program was written in Fortran 90. The DOS and Unix executable programs, the user's guide, and some example files are freely available for research purposes at http://www.bdporc.irta.es/estudis.jsp. The main feature of the program is to compute Monte Carlo estimates of marginal posterior distributions of parameters of interest. The program is quite flexible, allowing the user to fit a variety of linear models at the level of the mean and the logvariance.