Changes in spatio-temporal localization of tripeptidyl peptidase II (TPPII) in murine colon adenocarcinoma cells during aggresome formation: a microscopy study based on a novel fluorescent proteasome inhibitor.

Extralysosomal proteolysis is a multistep process involving the Ubiquitin- Proteasome System (UPS) and supplementary peptidases. Tripeptidyl peptidase II (TPPII) is the most extensively characterized enzyme, supplementing and sometimes substituting for proteasomal functions. In response to proteasome inhibition, polyubiquitinated proteins acting as proteasome substrates aggregate with proteasomes and form aggresomes. Several proteasome inhibitors are used as anti-cancer drugs. Thus, in our study, we used a novel fluorescent-tagged proteasome inhibitor BSc2118 to induce aggresome formation in C26 murine colon adenocarcinoma cells. It allowed us to obtain effective, inhibitor-based, proteasome staining in vivo. This method has been validated by standard post-fixed indirect immunostaining and also allowed co-immunodetection of TPPII and polyubiquitinated proteins under laser scanning confocal microscopy. We found that in the absence of the inhibitor, TPPII is diffusely dispersed within the cytoplasm of C26 cells. The proteasome and ubiquitin-rich perinuclear region failed to display enhanced TPPII staining. However, when proteasome function was impaired by the inhibitor, TPPII associated more closely with both the proteasome and polyubiquitinated proteins via TPPII recruitment to the perinuclear region and subsequently into emerging aggresomal structures. Furthermore, we have demonstrated the dynamic recruitment of TPPII into the developing aggresome: TPPII in the early aggresome was dispersed within the central part but subsequently aggregated on the surface of this structure. In the mature aggresome of C26 cells TPPII formed a spherical mantle, which surrounded the round core containing proteasomes and polyubiquitinated proteins. Our morphological data indicate that TPPII displays spatial localization with proteasomes especially upon proteasome inhibition in aggresomes of C26 cells.

[Patient's course requiring NIPPV in Centre Hospitalier Lyon-Sud]. / Le parcours du patient nécessitant une VNI au centre hospitalier Lyon-Sud.

Non-invasive positive pressure ventilation (NIPPV) has become a major therapeutic of acute respiratory failure. Thanks to technical progress, its use has become widespread in intensive care units and now in emergency and pneumology departments, for indications recognized and validated as decompensation of chronic obstructive pulmonary disease and acute cardiogenic pulmonary edema. Patients with this conditions transit in the hospital, from the emergency or pulmonology departments, sometimes through intensive care units. Knowledge of the NIPPV, its indications, contraindications, terms of use and surveillance requires trained teams. This training covers not only the technical but also the hardware, multiple ventilation modes, and interfaces. Other indications being evaluated, such as ventilation in the perioperative period, also require coordination between different actors. The establishment of a specific group of thinking and working around the NIPPV is clearly needed, allowing teams of hospital (emergency department, intensive care unit, pulmonology, anesthesia) to work together. This work deals with different areas: training, equipment, condition of receiving patients in the different services within the constraints of personnel and equipment. In this article, we trace the point of view of each of the professionals in this group and some of the actions implemented.

[Multicenter evaluation of h-FABP semi-quantitative assay (Cardio Detect) in central laboratory: the point in acute myocardial infarction diagnosis]. / Evaluation multicentrique du dosage semi-quantitatif de la h-FABP (Cardio Detect) au laboratoire central : intérêt dans le diagnostic de l'infarctus du myocarde.

The diagnostic performance of heart-Fatty Acid Binding Protein (h-FABP) (semi-quantitative CardioDetect test) and cardiac troponin I (TnIc) blood assays were compared in one hundred patients presenting with suspicion of acute coronary syndrome. Final patient diagnosis was "acute myocardial infarction" in 36 cases, "non ST myocardial infarction" in 25 cases and "non ischemic pathologies" in 39 cases. h-FABP results were positive in 26 patients, negative in 57 patients and ambiguous in 17 patients, the latter corresponding to the final diagnosis of "acute myocardial infarction" in 5 cases, "non ST myocardial infarction" in 2 cases and "non ischemic pathologies " in 10 cases. At admission, h-FABP and TnIc exhibiteda sensitivity of 54% an 66%, respectively and a specificity of 86% and 95%, respectively. Positive and negative predictive values were 81% and 64% for h-FABP, respectively and 92% and 75% for cTnI, respectively. h-FABP and cTnI demonstrated a similar diagnostic efficiency if admission delay is less than 4 hours after onset of chest pain (area under ROC curve TnIc = 0.767 +/- 0.091 ; area under ROC curve h-FABP = 0.622 +/- 0.109 ; p = 0.144). On the contrary, cTnI assay demonstrated a better efficiency than h-FABP (p< 0.005) for patients admitted in a delay of 4 to 12 hours after the onset of chest pain. If chosen cTnI cut-off corresponded to the recent consensus definition used for monitoring acute coronary syndrome patients, h-FABP semi-quantitative assay realized within central laboratory did not demonstrated a better diagnostic efficiency than cTnI.

Infection characteristics of rabies virus variants with deletion or insertion in the pseudogene sequence.

We investigated the infection characteristics of recombinant rabies virus variants modified in the pseudogene sequence. Infection of neuronal cell lines by the SAD W9 and SAD V* variants (respectively with deletion or insertion in this sequence) showed no significant differences as compared to the parental strain, the attenuated strain SAD B19, in infection characteristics such as number of infected cells or viral yield. The inoculation of mice by these variants resulted in similar infection patterns and pathogenicity. Stereotaxic inoculation of the different variants into the rat striatum showed that deletion or insertion did not affect the axonal virus spread, nor did insertion of a complete additional transcription unit, that could be expressed in the areas connected to the inoculation site. These results show that the pseudogene sequence is not involved in viral spread and pathogenicity and confirm the availability of this domain for targeting and expression of foreign genes into neurons.

Enquête éco-pathologique continue: facteurs de risque des mammites de la vache laitière I. Analyses multidimensionnelles sur données d'élevage.

An epidemiological study about mastitis in French dairy herds, supported by an ecopathological survey is described.The aim of this work was to explain variations of the annual frequencies of clinical mastitis in the farms studied and of milk somatic cell counts as predictive of the infectious status of the mammary gland. Milk cell counts were expressed as annual percentages of cell results < 2.10(5) cells / mL (0.2 CEL) and > 1.10(6) cells / mL (1.0 CEL) in each herd.Twenty-nine herds were studied from June 1979 to July 1981 (22 to 92 cows per herd, Normande and Pie-Noire breeds) and, for the first time, the influence of 88 variables upon clinical mastitis, 0.2 CEL and 1.0 CEL was performed by multidimensional methods.These analyses led to the evidence of two kinds of factors linked with mastitis occurrences or milk cell-counts:1) Not well defined factors: geoclimatic, racial, links between production traits, extra-mammary pathology, susceptibility to mastitis and cells-counts; to elucidate if they were factors really linked with mammary infections, a complementary analysis is described as necessary (cf. second part of this study).2) Risk or protective factors, which appeared as: - absence of at least one annual examination of the milking machine (risk factor for clinical mastitis) - udder washing with individual towels, associated with enough straw for bedding (protective factor against mastitis) - teat dipping (associated with low milk cell-counts)Other prophylactic methods appeared without any effect. In the discussion, main difficulties of an epidemiological approach to the mastitis problem are summarized. A convenient hygiene-prophylaxis interaction is necessary to control mammary infections in dairy herds.

[Continuing eco-pathological survey: 6. Influence of zootechnical and space-time factors on some biochemical variables in the dairy cow]. / Enquête éco-pathologique continue: 6. Influence de facteurs zootechniques et spatio-temporels sur quelques variables biochimiques chez les vaches laitières.

Several hundred of blood samplings were performed on dairy cows in four French departments, from 1978 to 1982. They were used to analyse the effect of eight factors on the physiological variations of twelve biochemical variables. Only the blood withdrawal date related to calving date or milk production were able to explain more than 15% of the variance of free or total cholesterol or phospholipids.

[Continuing eco-pathological survey: 10. Variation of pathological frequency in dairy cow stock as a function of season, year and number of lactations]. / Enquête éco-pathologique continue: 10. Variations des fréquences pathologiques en élevage bovin laitier en fonction de la saison, de l'année et du numéro de lactation.

The distribution of main herd diseases in course of time is studied with the season, the year and number of lactation. There is a seasonal effect for retained placenta, spring and summer being a favorable time for the increase of their frequency. Milk fever is also more frequent during summer. Out-calving pathologies are more frequent during housing time, but we do not observe any monthly variation for ovarian pathology. The age is a risk factor for milk fever and retained placenta. On the contrary, dystocia and stillbirth are more common on heifers. All the metabolic disorders and infectious diseases (mastitis-metritis) seem to be more frequent on old cows. The variations between years are interpretable with difficulty because they depend on the regularity in the quality of collected data. More, the necessity for the breeders to observe conscientiously their animals is a favouring factor for the improvement of the health status of the herd.

[Continuing eco-pathological survey: 8. Epidemiological approach to postpartum mastitis in the dairy cow: discriminating biochemical parameters]. / Enquête éco-pathologique continue: 8. Approche épidémiologique des mammites post-partum chez la vache laitière: paramètres biochimiques discriminants.

To discriminate two groups of dairy cows (with or without post-partum mastitis), with blood parameters, we used discriminant analysis on two samples of dairy cows: one sample where blood was collected around parturition (15 days before and 15 days after calving); one sample where blood was collected at any time of the lactation, but with the same mean interval calving-blood collection in the two groups of animals. Blood parameters which differenciate the two groups (ill and safe) are essentially parameters of energetic metabolism i.e. glucose and 3-hydroxybutyrate. Results are discussed according to the possibility of glucose to be an indicator of energetic status.

[Continuing eco-pathological survey: 11. Development of pathological frequency in dairy cow stock as a function of the stage of lactation]. / Enquête éco-pathologique continue: 11. Evolution des fréquences pathologiques en élevage bovin laitier en fonction du stade de lactation.

The study of disease distribution observed during lactation shows that three patterns of pathology may be described: diseases for which critical period is post-partum time: we observed during the first month of lactation 32.2% of cases of clinical mastitis, 43.9% of cases of metritis, 37.9% of digestive disorders, 59.6% of appetite troubles, 23.1% of treat lesions and 52.9% of udder edema. The monthly curve squares with asymptotic function. foot disorders for which the high milk yield production period is the more sensible time (three first month of lactation). The monthly curve, near a linear decreasing, squares with a quadratic or square-root function. ovarian pathology for which frequencies depends more acute attention from breeders or clinicians to reproductive disorders than objective observation of ovarian dysfunction.

[Continuing eco-pathological survey: 4. Demonstration of pathologic associations in dairy cow stock: herd data]. / Enquête éco-pathologique continue: 4. Mise en évidence des associations pathologiques en élevage bovin laitier: données d'élevage.

Data concerning frequencies of 17 main diseases in 59 dairy farms from continual eco-pathological survey, are analyzed by classical regression test, principal components analysis and analysis of variance. Thirty-three statistical relations (positive simple correlations) are shown. Clinical mastitis and retained placenta are associated to nine diseases each, metritis, stillbirth and non-infectious foot disorders to six, dystocia and infectious foot disorders to four, pathology of calf and abortion to three, mammary edema and appetite disorders to two. Ovarian pathology, digestive disorders and teat lesions are "isolated", being associated to only one other disease. The pathological frequencies in dairy farms seem to change in the same way, indicating the existence of common risk factors, and leading to propose a global hygienic prevention. The pathological associations allow to definite some groups of dairy farms showing one or other dominant pathological complex. In the frame of our sample of dairy farms, two groups seem to be determined: one with calving disorders and calf pathology centred on retained placenta, the other with foot disorders, infectious and metabolic troubles, centred on mastitis, as dominant diseases.

[Continuing eco-pathological survey: 5. Demonstration of pathologic associations in dairy cow stock: individual data]. / Enquête éco-pathologique continue: 5. Mise en évidence des associations pathologiques en élevage bovin laitier: données individuelles.

The calculation of pathological association ratio (or relative risk) from two ways contingency tables, allowed to show numerous interrelationships between diseases of 1,205 dairy cows. The analysis concerns the animal during productive life, the lactation (all lactation mixed), a particular one (1st, 2nd, 3rd and more) and the beginning of productive life of cow (three first lactations). The relationships are discussed according to the level of calculation (animal, lactation, or beginning of life). There is some triangular relations: calving trilogy (dystocia-stillbirth-retained placenta), infectious trilogy (foul of the foot-metritis-mastitis), metabolic trilogy (appetite disorders-ketosis-digestive disorders), foot disorders trilogy (lameness-non infectious diseases of foot-foul of the foot). These pathological groups seem to depend from common risk factors (climate-feeding-cowshed hygiene-control of heat-age-culling policy-genetic factors-anatomical or physiological predisposition, animal size...), but, numerous interactions between groups are also shown, leading to consider pathology in farm like a complex entity.

[Continuous ecopathological survey. 1. Classification of the pathology observed in dairy cattle breeding]. / Enquête éco-pathologique continue: 1. Hiérarchie de la pathologie observée en élevage bovin laitier.

Observations were made during a continuous ecopathological survey on 3,216 dairy cows from 90 dairy farms and localised in six French departments, over a three year period (1978-1981). The pathology described for cows and their calves was classified according to the frequency of each disease and/or symptom. The reasons for culling as well as those for mortality were also classified. The pathology of reproduction and the mammary gland in the dairy cow and the digestive pathology and mortinatality of calves appear to be the major pathological problems.

[Continuous ecopathological survey. 2. Typology of dairy cattle farms with high and low fertility]. / Enquête éco-pathologique continue: 2. Typologie d'élevages de vaches laitières à bonne et à mauvaise fertilité.

Management characteristics were analysed in two groups of dairy herds (20 to 60 cows per herd) during a continuous eco-pathological survey, in relation to high and low fertility. 59.8% of the cows from the eleven high fertility herds were successfully inseminated at first service and 11.4% of the cows inseminated for the first time more than 40 days post-partum required 3 or more services. The corresponding values were 51.1 and 25.6% for the ten low fertility herds. Other reproductive criteria (calving-first service interval, disturbed oestrus cycles), which more depend on the reproduction management, were not different between the two groups. Feeding system greatly varied in the low fertility group. Frequent changes in diet composition, a longer period of "green" forages distribution (crucifers, grass silage) as well as less utilisation of concentrates are some of the reasons which can explain the low fertility. Calving occurred mainly from February to April in low fertility herds, at the transition from winter diets to grazing. Milk production was lower, 4 580 kg per cow and per year vs 5 520 kg for high fertility group. Milk production falls were more frequent and Holstein semen was less used. Tied housing conditions, and higher incidence of Fasciola hepatica and abomasal mematodes characterised low fertility herds; inversely, the clinical pathology appeared to be less frequent in this group. Soil manganese concentration was lower (16.3 ppm) in the low fertility group as compared to the other group (30.9 ppm); this could be explained by less schistose rocks where the low fertility farms are localized. In conclusion, the risk factors associated with low fertility may be attributed to: the composite system of feeding in addition to the use of "green" forages, a lack of proper programming with regards to the season of calving, and the high frequency of parasitic infestations, with special reference to liver fluke.

[The control of animal diseases in france during the last fifteen years, as it appears throught the eradication of bovine tuberculosis. proposals for the future (author's transl)]. / La lutte contre les maladies animales en France au cours des 15 dernieres années vue au travers de la prophylaxie de la tuberculose bovine. Propositions pour l'avenir.

In the general outline of the success, on a national scale, of the eradication of bovine tuberculosis in France up to 1974, we have tried to find out the factors which could explain the differences, sometimes important, between French departments in the efficiency of this eradication. We have selected twenty seven parameters distinctive of each of the 74 departments having more than 50 000 bovine under the control of State Veterinary service. These parameters describe 1) tuberculosis infection rates of animals and herds, at three periods; 2) some structural characteristics of bovine production; 3) sanitary environment (veterinary practicioners, State Veterinary service, breeders association for sanitary protection). Statistical analysis by principal components show that department infection rate decreases as the first three factors evidenced by this analysis vary positively; they are: 1) "Sanitary Consciousness", which is defined both by the important of bovine production and by precociousness in developing prophylactic measures, these two factors being independent. 2) Specialization in bovine production, characterized by the percentage of young animals reared in the herd and by the proportion of grass produced by ley. 3) Intensity of sanitary environment. These three factors, which are interlinked to various degrees, control respectively 25%, 13% and 13% of the total variance. A complementary analysis of the data suggest that the efficiency of eradication could be improved by a better knowledge of the movement of breeding animals between the departments, and by a more careful recording, in some instances, of infected animals in slaughter hourses. When the importance of a herd increase sanitation of tuberculosis in infected herds is less readily achieved and brucellosis abortion rate increases. At the departement level, efficiency of eradication is not correlated to the expenses of prophylaxy. For the development of an integrated sanitary policy, taking into account such non specific factors as those exemplified in the study could help to a better control of animal diseases, in parallel to the desirable improvement of the specific means of combatting each disease.

Prognosis of diarrhoea in the newborn calf: statistical analysis of blood biochemical data.

Blood samples were taken from 55 diarrhoeic calves (neo-natal diarrhoea) at the time of the first therapeutic intervention. Eleven blood parameters were measured in these samples. The whole group of measurements were analysed by their principal components (tables 3 and 4). The results are expressed in visual form by Figures 1 and 2. The first principal axis corresponds to the acid-base balance while the second axis can be considered as corresponding to the catabolism. The discriminant analysis (table 5) shows that the parameter having the best prognosis value is the blood urea concentration. By adding two other easily measured parameters (the haematocrit and the blood chloride concentration) the probability of classifying correctly the calves into one or other of the groups dead or surviving is approximately 80%.

Effect of intravenous infusion of glucose and/or fructose on the composition of blood plasma and the clinical response of the calf.

Intravenous infusions of glucose (0.4 g/kg) or glucose (0.4 g/kg) + fructose (usually 1 g/kg) to calves were followed by various clinical reactions from absence of reaction up to death of a few calves. Fructosemia. After fructose + glucose infusion, the blood fructose concentration reached a level inferior to the highest value observed after only glucose infusion, except in the calves dead after the perfusion and in the newly-born calves (the initial fructose concentration was not nil). Glycaemia. In any case, fructose associated with glucose, did not eliminate the rebound of hypoglycaemia following usually a perfusion of glucose. In calves where the death occurred, the hypoglycaemia was specially marked.

Level of linoleic acid in pericardial lipids of Charollais cows with and without ovarian cysts.

The content of linoleic acid, a precursor of prostaglandins, in pericardial lipid store is higher in cows with normal ovaries than in cows with cystic ovaries.

Reovirus-like agent (rotavirus) associated with neonatal calf gastroenteritis in France.

A reo-like virus (calf rotavirus) was shown to be associated with cases of neonatal calf diarrhea in France. The virus could be detected in more than 50 p. 100 of diarrheic fecal samples, while it was practically absent in control samples originating from healthy calves that had never had diarrhea. The results obtained by electron microscopy and immunofluorescent studies indicate that the virus is closely related or identical to the agent isolated originally in the United States.