[Admissions to a coronary care unit for severe iatrogenic conditions: a prospective study of 7244 patients]. / Les admissions pour iatrogénie sévère en unité de soins intensifs de cardiologie: étude prospective sur 7244 patients.

INTRODUCTION: Iatrogenic complications are defined as adverse drug reactions or complications induced by non-drug interventions, such as cardiac devices or stimulation techniques. Iatrogenic complications occurring during hospital stay are known to be associated with increased hospital length of stay and mortality. Only few data are available on iatrogenic as cause of hospital admission, particularly in coronary care unit. In patient admitted in coronary care unit for iatrogenic, we aimed: (a) to analyse their prevalence, type and characteristics, (b) to analyse their in-hospital length of stay and mortality and (c) to evaluate the predictive factors of severity and mortality. METHODS: From 1st April 2008 to 31 January 2012, all the consecutive admissions caused by iatrogenic complications at the coronary care unit were prospectively included and classified in two groups: (1) pharmacological iatrogenic (beta-blockers, digoxin, calcium channel blockers, cordarone, several antiarrhythmic , anticoagulants, antiplatelets and others), (2) non-pharmacological iatrogenic (pacemaker, cardioverter-defibrillator, radiofrequency, coronary angiography and cardiac surgery including valve surgery). We excluded patients with intentional overdose. We also compared patients according to the severity (group 1: patients who just need a monitoring; and group 2: patients for whom there was invasive procedure or for whom we used vasoactive amine). RESULTS: Among 7244 patients admitted in coronary care unit during the inclusion period, 250 (3.4%) were admitted for iatrogenic complication, 136 in pharmacological group and 114 in non-pharmacological group. In non-pharmacological group, there was more men: 73.7% vs. 47.8% (P < 0.001), patients are younger: 67.3 ± 13.2 vs. 75.4 ± 15.8 (P < 0.001) and are more severe: 80.4% in group 2 vs. 69.4% (P = 0.05). The mortality in this group tends to be more important. According to the severity, there is no difference about drugs: 7.4 ± 3.4 vs. 6.8 ± 2.9 (P = 0.184) and are staying longer in hospital: 4.7 ± 3.2 days vs. 3.4 ± 2.4 (P = 0.009) for coronary care unit length of stay and 15 ± 13.7 vs. 10 ± 9.8 (P = 0.003) for total length of stay. CONCLUSION: Iatrogenic represent a non-negligible cause of admission in coronary care unit, which associated with significant mortality (8.8%) and with a trend toward a higher length of stay. Further studies are needed to determinate the origin of mortality and to better characterize patients at risk of iatrogenic.

Construction and characterization of a conditionally active version of the serine/threonine kinase Akt.

Akt is a serine/threonine kinase that requires a functional phosphatidylinositol 3-kinase to be stimulated by insulin and other growth factors. When directed to membranes by the addition of a src myristoylation sequence, Akt becomes constitutively active. In the present study, a conditionally active version of Akt was constructed by fusing the Akt containing the myristoylation sequence to the hormone binding domain of a mutant murine estrogen receptor that selectively binds 4-hydroxytamoxifen. The chimeric protein was expressed in NIH3T3 cells and was shown to be stimulated by hormone treatment 17-fold after only a 20-min treatment. This hormone treatment also stimulated an approximate 3-fold increase in the phosphorylation of the chimeric protein and a shift in its migration on SDS gels. Activation of this conditionally active Akt resulted in the rapid stimulation of the 70-kDa S6 kinase. This conditionally active Akt was also found to rapidly stimulate in these cells the phosphorylation of properties of PHAS-I, a key protein in the regulation of protein synthesis. The conditionally active Akt, when expressed in 3T3-L1 adipocytes, was also stimulated, although its rate and extent of activation was less then in the NIH3T3 cells. Its stimulation was shown to be capable of inducing glucose uptake into adipocytes by stimulating translocation of the insulin-responsive glucose transporter GLUT4 to the plasma membrane.

[Iscom (Immuno Stimulating Complex)-vaccine of equine influenza virus--transmission electron microscopic investigation and literature review]. / Iscom (Immuno Stimulating Complex)-Vakzine des equinen Influenza-Virus--transmissionselektronenmikroskopische Chargenüberwachung und Literaturübersicht.

Vaccines on the base of immunostimulating complexes are very rare in human and veterinary practice. Until now, Iscom vaccines mainly have been developed for scientific experimental investigations. Synthesis and preparation of Iscom vaccines, mainly basing on the reviewed literature, as well as electron microscopical investigations of Iscom structures and the monitoring of vaccine fractions of Iscoms are described. The equine influenza Iscom vaccine, developed by the Mallinckrodt Veterinary GmbH, is one of the first commercial Iscom vaccine used in veterinary medicine. In comparison with other commercially used vaccines, depending on the high level of antigen presentation of Iscom structures, a ten times higher antibody response is to be expected by the use of Iscom vaccines.

A nonradioactive assay for the insulin receptor tyrosine kinase: use in monitoring receptor kinase activity after activation of overexpressed protein kinase C alpha and high glucose treatment.

In the present study we describe a nonradioactive assay for measuring the intrinsic tyrosine kinase activity of the insulin receptor. This assay utilizes as an exogenous substrate a biotinylated peptide based on the sequence of the endogenous substrate insulin receptor substrate-1 (IRS-1). To separate the tyrosine phosphorylated peptide from the nonphosphorylated peptide, immobilized recombinantly produced SH2 domain of the p85 subunit of the phosphatidylinositol 3-kinase is utilized to bind the tyrosine-phosphorylated peptide. The amount of bound peptide is then detected by the use of peroxidase-conjugated streptavidin and a colorimetric assay. This assay has been used to measure the tyrosine kinase activity of receptor which was immunocaptured from lysates of various cells overexpressing the human insulin receptor as well as the endogenous insulin receptors in the parental cells. In this in vitro assay, no decrease in tyrosine kinase activity was observed in receptors from cells with activated overexpressed protein kinase C alpha or after high glucose treatment although a decrease in in situ phosphorylation of IRS-1 was observed with the activation of protein kinase C alpha. These results indicate that this assay may be a useful new method for monitoring the enzymatic activity of the insulin receptor kinase as well as other tyrosine kinases.

IGF-I and insulin receptors in bovine skeletal muscle: comparisons of different developmental ages, two different genotypes and various individual muscles.

To investigate the regulation of the IGF-I and the insulin receptor in bovine skeletal muscle, we determined their concentrations and their affinity constants in animals of different age, muscle type and breed. Receptors were solubilized and measured by an enzyme-immunoassay. During ontogenesis the concentration of IGF-I receptors decreased from 1650 /+- 600 fmol/mg protein in 3-5-month old fetuses to 105 /+- 20 fmol/mg protein in 16-month old heifers (p < or = 0.01). The insulin receptor concentrations also decreased with age from 136 /+- 28 fmol/mg protein in 3-5-month old fetuses to 50 /+- 11 fmol/mg protein (p < or = 0.05) in 16-month old heifers. In order to compare different muscle types, seven muscles, which represent large differences in fibre type composition and growth impetus, were selected from 6 month old female Jersey calves and were assayed for IGF-I and insulin receptors. We observed significant differences of the IGF-I as well as the insulin receptor concentrations between distinct muscles. However, no relationship could be established between receptor concentration and fibre type composition or growth impetus. In muscle of two cattle breeds, differing markedly with regard to muscle growth intensity, the Jersey and the German Fleckvieh breed, we observed no divergence in IGF-I nor insulin receptor concentrations. We found no differences in IGF-I and in insulin receptor affinities in any of the adult animals.(ABSTRACT TRUNCATED AT 250 WORDS)

Molecular polymorphism as a tool for differentiating ground beetles (Carabus species): application of ubiquitin PCR/SSCP analysis.

Differentiation between Carabus species (ground beetle) and subspecies is difficult, although there have been extensive studies. To address this problem we have applied PCR in combination with SSCP analysis focussing on the evolutionally conservative ubiquitin gene to elaborate a new approach to molecular differentiation between species. We report that Carabidae possess an ubiquitin gene and that its gene has a multimeric structure. Differential SSCP analysis was performed with the monomeric form of the gene to generate a clear SSCP pattern. Such PCR/SSCP resulted in reproducible patterns throughout our experiments. Comparing different Carabus species (Carabus granulatus, C. irregularis, C. violaceus and C. auronitens) we could observe clear interspecies differences but no differences between genders. Some species showed some remarkable differences between the individuals. We suggest that the ubiquitin PCR-SSCP technique might be an additional tool for the differentiation of ground beetles.

An enzyme immunoreceptor assay for the quantitation of insulin-like growth factor-I and insulin receptors in bovine muscle tissue.

To investigate the regulation of insulin-like growth factor-I (IGF-I) and insulin receptor in muscle, a sensitive enzyme immunoreceptor assay (EIRA), which allows for the determination of both affinity and capacity of the receptors, was developed. After solubilization with Triton X-100, receptors were immobilized in microtiter plates using receptor specific monoclonal antibodies that recognize the intracellular beta-domain of the respective receptors (clone 17A3 and clone 29B4). The immobilized receptors were labeled with either biotinylated IGF-I or insulin. The bound hormones were detected with a streptavidin-horseradish peroxidase technique. The assay had a detection limit of 1 fmol receptor/well. The intraassay variation was 9% (n = 22) for the IGF-I receptor concentration and 12% (n = 22) for the insulin receptor. The interassay variation was 5% (n = 4) for the IGF-I receptor and 10% (n = 4) for the insulin receptor. The coefficient of variation of the dissociation constants (Kd) was 26% (n = 7) for the IGF-I receptor and 24% (n = 7) for the insulin receptor. The assay system was used to study the effect of growth hormone treatment upon IGF-I and insulin receptors in bovine skeletal muscle. Three groups of 12 heifers (13 months old) were treated with either 320 or 640 mg recombinant bovine somatotropin (slow release preparation) every fortnight for 3 months. When samples of m. splenius were assayed for IGF-I and insulin receptors, there was no difference between groups for receptor concentration or affinity.(ABSTRACT TRUNCATED AT 250 WORDS)

Action of metal chelators on lipoxygenases, cyclooxygenase and on inflammation-induced vasodepression.

Metal complexing agents could exert antiinflammatory activity by inhibition of oxygen radical production, by inhibition of eicosanoid synthesis and by inhibition of metalloenzymes. We found a moderate antiedema activity of the iron (II)/iron (III)-chelator combination, o-phenanthroline/desferrioxamine, which could refer to an involvement of iron ions in the inflammatory reaction. The newly synthetized complexing agent ethylene-diimino-dibutyric acid caused weak antiedema and antivasodepressor effects which remain to be explained. Altogether, the in vivo effects of the metal chelators were rather weak. In vitro, only desferrioxamine produced a remarkable inhibition of two lipoxygenases.

Guinea pig protection test as indicator of potency of oil emulsion foot-and-mouth disease vaccines.

A vaccine potency test is described involving virus challenge to six groups of 10 guinea pigs at five weeks after vaccination. Sixteen oil emulsion foot-and-mouth disease vaccines were so tested and nine retested after storage at 4 degrees C for up to 28.3 months. The results were compared with those of the routinely used oil emulsion vaccine potency test (protection afforded to eight pigs challenged 21 days after vaccination). When guinea pig estimates of 3 log2 PD50 or more were obtained, then, with one exception, the batches protected all or almost all pigs from challenge, but when the guinea pig estimates were less than 1 log2 PD50, the vaccines failed to protect five out of eight pigs. The sensitivity and reproducibility of the guinea pig method, established by repeated tests on two vaccine batches, seemed acceptable. The results suggested that guinea pig estimates might provide a suitable substitute for pig challenge potency tests because they reflected the potency of the vaccines, were likely to involve smaller standard errors and caused less discomfort to animals.

Production of rabies vaccine by an industrial scale BHK 21 suspension cell culture process.

A process for the production of an inactivated rabies vaccine for animal use is described, whereby the LEP Flury strain of rabies virus is grown in 1 000 L cultures of BHK 21 suspension-adapted cells. The virus is inactivated with aziridine and, after concentration and purification by hollow fibre ultrafiltration, is adsorbed onto aluminum hydroxide gel. The BHK 21 suspension-adapted Master Cell Seed used has satisfied all the requirements of the US Code of Federal Regulations, Title 9, (1977) for cell lines. Cultures prepared from it produce high yields of rabies virus antigen and vaccines with an average potency of 6 IU can be produced without difficulty. Such vaccines give at least 3 years' protection in dogs and are safe to use in all species. Experience gained with the closely analogous process for the production of FMD vaccines suggests that the process can be readily transposed and successfully operated in developing countries if and when it becomes cost-effective to do so.

The relationship between serum antibody titres and protection from foot and mouth disease in pigs after oil emulsion vaccination.

In trials performed in Brazil, England and Germany, 765 pigs were challenged at one, three to four or 17 weeks after vaccination and their sera were titrated by macrocolour, microcolour or microcytopathic effects neutralization tests. Using O1 virus strains and challenge intervals of three to four weeks, significant correlations (P less than 0.01) were demonstrated between protection and serum neutralizing titres although the titres corresponding to given levels of protection varied with the titration method employed. The protective capacity of antiserum was confirmed by passive antibody transfer experiments. Retrospective analysis of 49 challenge tests showed that when the macrocolour test was employed, a group mean (n = 8) serum titre of 1.74 log10 SN50 corresponded with five out of eight (or more) protected from challenge in 92% of cases. The possibility of using serum titres as an alternative to challenge for oil emulsion vaccine potency tests is discussed.

Revaccination response of cattle as a function of the 140S foot and mouth disease antigen dose.

Fifty adult cattle were subdivided in 2 equal groups and inoculated with foot and mouth disease vaccines containing either 5 micrograms or 0.01 microgram of O1BFS 1860 140S antigen per 3 ml dose. Four months later they were subdivided into groups of 5 and injected with vaccines containing concentrations of the 140S antigen ranging from 54 micrograms to 7 ng per 3 ml dose. Bleedings were carried out periodically and the serum neutralizing antibody titres showed that the revaccination responses were influenced by the dose of 140S antigen in both the primary and secondary vaccinations. A linear log relationship was demonstrated between the anamnestic serum antibody log response and the dose of antigen in the secondary vaccines.

The relationship between the 140S antigen dose in aqueous foot-and-mouth disease vaccines and the serum antibody response of cattle.

Groups of eight or 12 cattle were injected with dilutions of aqueous saponized vaccines containing 329 760 to 7 ng of 140S O1BFS 1860 strain foot-and-mouth disease antigen per dose. Four months later the animals were subdivided into groups of four and revaccinated with vaccines containing 329 760, 9160 and 42 ng of antigen per dose. Responses were established by taking serum samples from the animals periodically and examining these for neutralizing antibody activity. The results showed that after primary vaccination the dose of antigen in vaccines influenced the earliest time of protection, the peak antibody titres attained and probably, the duration of immunity. At 21 days after vaccination the log antigen-antibody response slope was linear between 7 and 9, 160 ng, with an inclination of 0.53, indicating that every doubling of the vaccine antigen dose produced an increase of 0.16 log10SN50 in the mean serum antibody titre. The secondary response was influenced by the antigen dose in the second vaccination and to a lesser extent by that in the first vaccination also.