The application of humane slaughterhouse practices to large-scale culling.

Mass culling is a strategy used inthe management of infectious disease outbreaks. The risk of disease spread due to animal transportation often precludes the use of slaughterhouses for this purpose, though they have been used on occasion. Consequently, culling takes place in the less-than-standardised environment on the farm. Regardless of where the cull occurs, the methods chosen to handle and kill the animals must preserve their welfare. This paper attempts to apply the best welfare practices from the controlled environment of the slaughterhouse to the mass culling of ruminants, pigs and poultry. It investigates the key welfare challenges and identifies astute planning executed by competent personnel as a crucial success factor. The urgency; capacity; species, type and age of the animal; personnel; and availability of equipment determine the restraint and killing methods used. The use of good on-farm restraining facilities and mobile killing devices can reasonably be expected to maintain welfare standards for ruminants and poultry. The pig's anatomy, natural behaviour, wide age range and housing types present additional challenges. Objective monitoring throughout the operation is vital for implementing immediate corrective measures whilst also informing procedural reviews. The authors suggest a monitoring tool based on a system currently used in abattoirs, but its limitations must first be validated.

Invasive aspergillosis in patients with acquired immunodeficiency syndrome: report of 33 cases. French Cooperative Study Group on Aspergillosis in AIDS.

PURPOSE: Acquired immunodeficiency syndrome (AIDS)-associated invasive aspergillosis (IA) is a rare condition, which is mainly reported as isolated cases either antemortem or at autopsy. The role of AIDS itself is controversial, because many of the reported patients exhibited the classic risk factors such as neutropenia and steroid therapy. The aims of this study were to report 33 patients with IA during AIDS and their outcome, focusing on the risk factors and the value of diagnostic procedures. PATIENTS AND METHODS: Thirty-three patients from 17 different medical centers in France were retrospectively included in the study. For pulmonary IA, we defined two types of patients: those with "confirmed IA," describing all the patients with histologically proven disease, and those with "probable IA," who had the development of a new pulmonary infiltrate on chest radiograph and a positive bronchoalveolar lavage (BAL) fluid culture for Aspergillus species without identification of other pathogens. For extrapulmonary IA, the diagnostic criteria included both positive histology and culture. RESULTS: Of the 33 cases included in this series, 91% were recorded during the last 3 years (1989 to 1991), suggesting that aspergillosis is an emerging complication in AIDS. Approximately 50% of the patients did not exhibit any classic risk factor, i.e., neutropenia and steroid treatment; almost all patients had a CD4 cell count less than 50/mm3. The mycologic culture from BAL was the method of choice for the diagnosis of invasive pulmonary disease because it was known to correlate well with histologic findings obtained either antemortem or postmortem. Of 28 patients with a positive BAL culture for Aspergillus, 15 underwent a biopsy or autopsy and 14 were positive at histology. Serum antigen detection was positive in only 4 of 16 tested patients. Clinical and radiologic signs did not differ from those observed in neutropenic patients without human immunodeficiency virus, except for the higher incidence of neurologic complications in AIDS. Interestingly, we observed three cases of invasive necrotizing tracheobronchial aspergillosis with acute dyspnea and wheezing. The use of amphotericin B (0.5 mg/kg/d) and/or itraconazole (200 to 600 mg/d) was most often unsuccessful. Only four patients experienced clinical and radiologic improvement. The mean interval between the diagnosis of IA and death was 8 weeks (range: 3 days to 13 months). CONCLUSIONS: This study suggests that aspergillosis is an important life-threatening condition in the advanced stage of AIDS. It requires an early diagnosis with BAL fluid culture and careful therapeutic evaluation.

The transport of animals in the European Union: the legislation, its enforcement and future evolutions.

Animal transport is an important activity of the farming industry. It is also the most controversial area of animal welfare. After presenting the current legislative framework in the European Union that protect animals during transport and the main enforcement difficulties, this paper considers the international, political and scientific perspectives that lead the European Commission to draft new rules in this regard.

[Acquired autoantibodies directed against human factor VIIIc. Treatment of 2 hemorrhagic accidents with porcine factor VIIIc]. / Auto-anticorps acquis contre le facteur VIIIc humain. Traitement de deux accidents hémorragiques par le facteur VIIIc porcin.

The emergence of an autoantibody directed against factor VIII may be responsible for severe, life-threatening haemorrhages. This rare disease is usually idiopathic, but it may be consecutive to an autoimmune disease or to the absorption of certain drugs such as penicillin. The diagnosis rests on the finding of a prolonged activated thromboplastin time with presence of a circulating anticoagulant and deep fall in factor VIII level. Two cases of severe haemorrhage successfully treated with porcine factor VIIIc are reported. The first case concerned an 80-year old woman presenting with a large haematoma of the thigh uncontrolled by injections of human factor VIIIc. The second case was that of a 24-year old woman in a state of shock due to a pleural blood effusion that occurred during heparin treatment of cerebral thrombophlebitis, combined with penicillin treatment of bronchial superinfection. In both cases the high-titer autoantibody to the human factor VIIIc did not, or little, cross with porcine factor VIIIc. Factor VIII rose after the first injection of the porcine factor, and the haemorrhage was rapidly controlled. In both cases, the autoantibody disappeared within a few months, either spontaneously or after treatment with immunosuppressants.

[Overload histiocytosis in 2 hemodialyzed patients. Possible role of dextran 40]. / Histiocytose de surcharge chez deux hémodialysés. Rôle possible du dextran 40.

The authors report two cases of singular overloading histiocytosis, observed in patients treated with regular dialysis and undergoing feverish cachectic disease. The observations of the two patients show a same histiocytosis infiltration of ganglions, liver, muscles, lung, pleura and pericardium, consisting in an accumulation of PAS positive intracytoplasmic substance, refringent at polarisation, microcrystalline on electron microscopy and on ionic analysis. Clinical likeness appears with PVP thesaurismosis but the histochemical findings are different. A favorable issue occurred with corticosteroïd treatment. Nature of PAS positive substance is uncertain. An exogenous origin is undoubted, and some elements call for the responsibility of dextran 40.

Persistence of ribosomal protein synthesis after infection of HeLa cells by herpes simplex virus type 1.

Because synthesis of rRNA persists late during herpes simplex type 1 infection and because S6 phosphorylation is always correlated with efficient translation of ribosomal protein mRNA, we tested the hypothesis that ribosomal protein synthesis and ribosome biogenesis could persist after infection. At different times after infection, proteins were labelled with 35S for 1 h before harvesting and ribosomes were purified. Measurement of radioactivity incorporated into individual ribosomal proteins separated by two-dimensional PAGE demonstrated that ribosomal proteins are still synthesized and assembled into mature ribosomes up to late times during infection, while synthesis of beta-actin is severely inhibited. During expression of late genes, ribosome biogenesis was estimated to be 58% of that of the control as judged by [3H]uridine incorporation into rRNA. As for beta-actin mRNA, the level of ribosomal protein mRNA decreased progressively from the beginning of infection, reaching about 30% of the control level during expression of late genes. Taken together, these results demonstrate that ribosomal proteins are still synthesized up to the late time of infection and efficiently assembled into mature ribosomes, while there is a severe shutoff of the synthesis of other cellular proteins.

Phosphorylation of herpes simplex virus type 1 Us11 protein is independent of viral genome expression.

The Us11 protein is a true late gene product of herpes simplex virus type 1 (HSV-1), whose exact function is unknown but which exhibits RNA-binding properties and which is phosphorylated on serine residues. In order to determine whether the Us11 protein is phosphorylated by cellular kinase(s) or by virally encoded kinase(s), the Us11 gene has been cloned and transiently expressed in HeLa cells. In addition, HeLa-derived cell lines have been selected for their ability to express Us11 protein constitutively. 32P-Labeling and analysis by two-dimensional electrophoresis of transiently and constitutively expressed Us11 protein demonstrated that, indeed, multiple phosphorylation of the protein occurs in absence of HSV-1 genome expression, indicating that the protein behaves as a natural substrate for cellular kinase(s). In addition, a sequence heterogeneity of the Us11 protein, due to a difference in the number of SPREPR repeats, has been characterized between different strains of HSV-1.

Phosphorylation of ribosomal protein L30 after herpes simplex virus type 1 infection.

In addition to an irreversible stimulation of S6 ribosomal protein phosphorylation, there is a modification of a subset of ribosomal proteins by phosphorylation after herpes simplex virus type 1 (HSV-1) infection. Moreover, in the course of this infection, three additional phosphorylated proteins can be extracted from ribosomes and separated by two-dimensional electrophoresis (2-DE) of total ribosomal proteins. One of them exhibits an identical molecular mass to L30, while being more acidic. This protein is phosphorylated on serine residues. The kinetics of appearance of this protein in the ribosomal fraction correlated with a decrease in L30 staining, as shown by 2-DE. Determination of the N-terminal amino acid sequence of this extra phosphoprotein and of L30-derived peptides demonstrated the identity of these two proteins.

The DNA sequence coding for the 5' untranslated region of herpes simplex virus type 1 ICP22 mRNA mediates a high level of gene expression.

The sequence coding for the 5' untranslated region (UTR) of ICP22 mRNA of herpes simplex virus type 1 has been tested for its ability to regulate gene expression. This sequence was placed in frame with the chloramphenicol acetyltransferase (CAT) coding sequence and under the control of the simian virus 40 early promoter-enhancer. Under these conditions, the sequence coding for the 5'UTR led to an increase of about 13-fold in CAT activity, measured during transient expression. The use of mutants with progressive deletions within the sequence coding for the 5'UTR allowed localization of the sequence responsible for the enhancement of gene expression to the first exon of the ICP22 gene. Precise quantification of hybrid ICP22-CAT mRNA showed that the sequence coding for the 5'UTR induced an increase in the amounts of transcripts, which resulted in a parallel increase in CAT activity. This increase in the level of hybrid ICP22-CAT mRNA is not the result of an increase in mRNA stability, nor is it due to more efficient nucleo-cytoplasmic transport of the transcripts. Moreover, the distribution of hybrid mRNA in the different ribosomal populations indicates that the 5'UTR of ICP22 mRNA does not induce a preferential recruitment of the transcripts by the translational apparatus. Taken together, these results indicate that a cis-acting element located in the sequence coding for the 5'UTR of ICP22 mRNA can mediate a high level of gene expression independently of the viral promoter and of viral trans-acting factors.

The herpes simplex virus type 1 US11 gene product is a phosphorylated protein found to be non-specifically associated with both ribosomal subunits.

Microsequencing of a cyanogen bromide peptide obtained from a basic phosphoprotein co-sedimenting with purified ribosomes extracted from herpes simplex virus type 1-infected human epidermoid carcinoma 2 cells identified this protein as a product of the true late US11 gene. An antibody was raised against a recombinant fusion protein expressed in Escherichia coli from a plasmid carrying 75% of the US11 coding sequence including the carboxy terminus. This antibody was used to probe Western blots carried out under various conditions of one- and two-dimensional electrophoresis. The electrophoretic behaviour of the immunoreactive proteins offered further proof that they were indeed products of the US11 gene. This US11 protein, which has phosphates on multiple serine residues, is brought into the cell by the virion and found to be present within ribosome fractions early after infection. This association with ribosomes is non-specific and due to probable aggregation or oligomerization of this proline-rich basic protein allowing its co-sedimentation with ribosomes during the different subcellular fractionation steps used for the purification of ribosomal subunits.

Cdk1 is a marker of proliferation in human lymphoid cells.

To better understand the relationship between the proliferation of human lymphoid cells and the expression of cdk1, a catalytic subunit of the histone H1 kinase (H1K), we examined its mRNA and protein content in 3 B-cell lines: Ramos, Reh-6 and IARC 963. Cells were elutriated according to their position in the cell cycle. Cell fractions were analyzed for cdk1 mRNA and protein cellular content by Northern blot and immunoblot, respectively, as well as for H1K activity. Both mRNA and protein amounts and H1K activity varied according to cell cycle phase, the lowest values being observed in G1-enriched fractions. For comparison, elutriated fractions were also tested for the expression of cdk2 and cdk4 proteins. Both showed some variations among fractions, but they were less clear than those of cdk1. We also tested 29 samples of lymphoid neoplastic and non-neoplastic tissues for proliferative activity (percentage of S and G2/M cells estimated by flow cytometry) and expression of cdk1, cdk2 and cdk4 proteins. We found a significant correlation between the percentage of cells in S or S + G2/M phases and cdk1 protein content but not cdk2 or cdk4 content. We conclude that cdk1 expression in human lymphoid cells varies during the cell cycle at both mRNA and protein levels.

Post-transcriptional transactivation of human retroviral envelope glycoprotein expression by herpes simplex virus Us11 protein.

Herpes simplex virus type 1 (HSV-1) Us11 protein, a true late gene product packaged within the virion, is delivered into cells after infection, exhibits a nucleocytoplasmic localization at early times, and later accumulates in the nucleoli. This RNA-binding basic phosphoprotein, capable of oligomerization, is supposed to be involved in post-transcriptional regulation of gene expression after HSV-1 infection. Expression of human T-cell leukaemia/lymphoma virus type-I (HTLV-I) and of human immunodeficiency virus type 1 (HIV-1) is post-transcriptionally regulated by Rex and Rev, respectively. These proteins are required for the cytoplasmic expression of unspliced gag-pol and singly spliced env transcripts. Here we show that HSV-1 Us11 protein is able to bind Rex- and Rev-responsive elements and to transactivate envelope retroviral glycoprotein expression.

[Torsades de pointes during sultopride poisoning]. / Torsades de pointes lors d'une intoxication par sultopride.

A case of sultopride poisoning (ingested dose 16 g) in a 35-year-old, 65 Kg man is described. On admission myoclonus, mydriasis, vomiting and cardio-respiratory arrest were observed. Torsades de pointes were treated with potassium chloride infusion and pace maker stimulation. Plasma sultopride concentration was 25 mg/l and urinary concentration 12 g/l. A prolongation of Q-T interval may announce severe arrhythmias in sultopride poisoning.