Detection of bacteriuria and bacteremia in newborn calves by a catalase-based urine test.

BACKGROUND: Bacteremia occurs frequently in newborn calves. The predictive value of clinical signs is low, suggesting the use of calf-side diagnostic tests. OBJECTIVES: To investigate testing of urine catalase activity (Uriscreen test) for bacteriuria and bacteremia detection. ANIMALS: Five colostrum-free calves and 3 colostrum-fed control calves. METHODS: Controlled experimental trial. Colostrum-free calves were inoculated PO with Escherichia coli O78+. A clinical score was established to define the onset of the illness. Blood and urine (cystocentesis) samplings and cultures, and Uriscreen tests, were performed 4-6 times from inoculation to death. Three control calves received the same management as 3 inoculated calves, but with colostrum and without inoculation. RESULTS: Bacteremia was demonstrated in all of the inoculated colostrum-free calves and in none of the control calves. The E. coli O78+ strain, E. coli, and Klebsiella spp. were recovered from 4/5, 5/5, and 2/5 inoculated colostrum-free calves, respectively. Urine cultures were negative for the 2 groups at the start of the experiment; 5/5 colostrum-deprived inoculated calves were positive for E. coli thereafter whereas 3/3 controls remained negative. Concordance of Uriscreen tests with bacteremia and bacteriuria was 0.86 and 0.88, respectively. Kappa value of agreement between Uriscreen and bacteremia and bacteriuria was 0.73 and 0.76, respectively. Sensitivity of Uriscreen for bacteremia and bacteriuria was 80.0 and 86.6%, respectively, and specificity was 92.8 and 88.8%, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: The results suggest that Uriscreen can be used for detection of bacteremia in neonatal calves in connection with a constant bacteriuria.

HPLC preparation of highly purified single-stranded M13 DNA.

Closed-circular, single-stranded viral DNAs are widely employed in DNA cloning and sequencing experiments. Because of their well-defined structure and sequence, closed-circular, single-stranded DNAs have also been used for ligand binding experiments and light scattering measurements. However, there is a high molecular weight impurity observed in light scattering experiments, which sometimes contaminates single-stranded DNA purified from phage that has been precipitated in polyethylene glycol, average molecular weight 8000, and purified by standard phenol-chloroform extraction. Three methods have been examined that remove this impurity from closed-circular, single-stranded M13mp19 DNA (SS M13 DNA). One employs a commercial peparation. This procedure yields pure but degraded SS M13 DNA, as shown by light scattering measurements and HPLC. Another employs a Whatman DE52 (diethylamino cellulose) column. This procedure yields intact DNA, but in poor yield (less than 20% of that obtained by phenol-chloroform extraction). The last was the most successful. This employs HPLC with a Waters AP-1 column with DEAE 8HR bedding. This procedure, which provides DNA in high yield (80%-90% column recovery) with an intact structure, is an efficient method for the isolation of high-purity, closed-circular, single-stranded viral DNA suitable for physical investigations and ligand binding measurements.

Intercalative DNA binding of model compounds derived from metabolites of 7,12-dimethylbenz[a]anthracene.

The DNA binding of nonreactive model compounds of metabolites of 7,12-dimethylbenz[a]-anthracene (DMBA)1 was studied in fluorescence quenching and fluorescence lifetime experiments. The model compounds examined were DMA and 8,9,10,11-tetrahydro-BA. DMA is a pi electron model of a highly carcinogenic bay region epoxide of DMBA, 8,9,10,11-tetrahydro-BA is a model compound of a less carcinogenic DMBA epoxide. The results indicate that the binding of DMA occurs primarily via intercalation. In 15% methanol the binding constant is 3.1 x 10(3) M-1. In 15% methanol and at DNA phosphate levels of 5.0 x 10(-4) M the intercalative binding of DMA is reduced by a factor of 6.2 when 5.0 x 10(-4) M Mg+2 is added. The DMA binding constant for intercalation is reduced by more than a factor of 4 when the methanol content of the solvent is increased from 0% to 20%. Finally DMA binding arising from pi interactions with the DNA bases is reduced more than 15 times when the DNA is denatured. For 8,9,10,11-tetrahydro-BA in 15% methanol the binding constant for intercalation is 6 times lower than that for DMA. These results along with previously reported binding data on other model compounds suggest that bay region metabolites of DMBA readily participate in physical pi stacking interactions with DNA.

UV photoelectron spectroscopy and ab initio characterization of valence orbital structures and conformations of neutral phosphate esters.

The HeI UV photoelectron spectrum of trimethyl phosphate (TMP) has been measured and interpreted with the aid of SCF molecular orbital calculations carried out with STO-3G, STO-3G* and 4-31G basis functions. The photoelectron spectrum of TMP is more accurately reproduced by results from 4-31G calculations than by results from STO-3G or STO-3G* calculations. However, all three basis sets yield results which predict the same assignment of the photoelectron spectrum. Results at the 4-31G level indicate that whether calculations are based on crystallographic bond angles and bond lengths or on STO-3G optimized geometries has little effect on the energetic ordering of the upper occupied orbitals. The energetic ordering of orbitals is also found to be only weakly dependent upon the torsional angle phi, describing rotation of ester groups about P-O bonds and upon the torsional angle psi, describing rotation of methyl groups about C-O bonds. For trimethyl phosphate, with C3 symmetry, the vertical ionization potentials of the upper occupied orbitals are 10.81 eV (8e), 11.4 eV (9a), 11.93 eV (7e), 12.6-12.9 eV (8a and 6e), 14.4 eV (7a) and 15.0-16.0 eV (5e and 6a). Calculations at the 4-31G level indicate that many of the highest occupied orbitals in neutral dimethyl phosphate and methyl phosphate have energies and electron distributions similar to orbitals in TMP. For TMP, a search for optimized values of phi and psi has been carried out at the STO-3G*level. In agreement with previous NMR studies and with classical potential calculations, the STO-3G* results indicate that both the gauche (phi = 53.1 degrees) and anticlinal (phi = 141.9 degrees) conformations are thermally accessible. Also in agreement with the classical potential calculations, the STO-3G* results predict that in the all gauche conformation energy is minimized when the methyl groups assume a staggered geometry (psi = 60 degrees to 80 degrees) and that an energy maximum occurs for an eclipsed geometry (phi = 0 degrees to 20 degrees). A study of the dependence of optimized values of O-P-O ester bond angles on the torsional angles, phi, was carried out at the STO-3G, STO-3G* and 4-31G levels. The results demonstrate that for C3 symmetry, the coupling of O-P-O angles to phi is influence by repulsive steric interactions.

A comparison of the intercalative binding of non-reactive benzo[a]pyrene metabolites and metabolite model compounds to DNA.

The reversible DNA physical binding of a series of non-reactive metabolites and metabolite model compounds derived from benzo[a]pyrene (BP) has been examined in UV absorption and in fluorescence emission and fluorescence lifetime studies. Members of this series have steric and pi electronic properties similar to the highly carcinogenic metabolite trans-7,8-dihydroxy-anti-9,10-epoxy-7,8,9,10-tetrahydrobenzo[a]pyrene (BPDE) and the less potent metabolite 4,5-epoxy-4,5-dihydrobenzo(a)pyrene (4,5-BPE). The molecules examined are trans-7,8-dihydroxy-7,8-dihydrobenzo[a]-pyrene (7,8-di(OH)H2BP), 7,8,9,10-tetrahydroxytetrahydrobenzo[a]pyrene (tetrol) 7,8,9,10-tetrahydrobenzo[a]pyrene (7,8,9,10-H4BP), pyrene, trans-4,5-dihydroxy-4,5-dihydrobenzo[a]pyrene (4,5-di(OH)H2BP) and 4,5-dihydrobenzo[a]pyrene (4,5-H2BP). In 15% methanol at 23 degrees C the intercalation binding constants of the molecules studied lie in the range 0.79-6.1 X 10(3) M-1. Of all the molecules examined the proximate carcinogen 7,8-di(OH)-H2BP is the best intercalating agent. The proximate carcinogen has a binding constant which in UV absorption studies is found to be 2.8-6.0 times greater than that of the other hydroxylated metabolites. Intercalation is the major mode of binding for 7,8-di(OH)H2BP and accounts for more than 95% of the total binding. Details concerning the specific role of physical bonding in BP carcinogenesis remain to be elucidated. However, the present studies demonstrate that the reversible binding constants for BP metabolites are of the same magnitude as reversible binding constants which arise from naturally occurring base-base hydrogen bonding and pi stacking interactions in DNA. Furthermore, previous autoradiographic studies indicate that in human skin fibroblasts incubated in BP, pooling of the unmetabolized hydrocarbons occurs at the nucleus. The high affinity of 7,8-di(OH)H2BP for DNA may play a role in similarly elevating in vivo nuclear concentrations of the non-reactive proximate carcinogen.

Effects of pre- or postpartum selenium supplementation on selenium status in beef cows and their calves.

The effect of Se supplementation before or after calving on Se status in deficient cows and their calves was studied using 72 beef cows in two experiments. In Exp. 1, cows calving in February or March 1997 were supplemented orally for 15 d in late pregnancy with 13.0, 32.5, or 45.5 mg of Se/d as sodium selenite. Glutathione peroxidase (GSH-Px) activities were measured in red blood cells (RBC) or plasma of cows and calves at d 15 and between d 17 and 88 after calving. In Exp. 2, cows calving in January 1997 were supplemented orally with .0, 13.0, or 32.5 mg of Se/d for 15 d postpartum, and calves were injected with 1.38 mg of Se when 2 d old and at an average age of 49 d. The GSH-Px activities were measured in 30-d-old calves and in cows and calves between d 77 and 115 after calving. In both experiments, Se supplementation resulted in adequate Se status for the dams. The increase in RBC GSH-Px activity was faster with 45.5 mg of Se/d, and GSH-Px activities remained high for up to 98 d after the end of supplementation. The improvement in Se status in calves as a result of maternal supplementation was greater in Exp. 1 than in Exp. 2, suggesting that the placental transfer of Se is more efficient than milk transfer. Prepartum oral Se supplementation of deficient beef cows with 13.0 mg of Se/d for 15 d allowed adequate Se status of dams and calves, and 45.5 mg of Se/d resulted in a faster improvement of Se status. Parenteral administration of 1.38 mg of Se to newborn calves did not sustain normal Se status in calves issued from deficient cows.

Effects of copper supplementation on the copper status of peripartum beef cows and their calves.

The effects of copper supplementation on the copper status of 40 late-pregnant Aubrac beef cows grazing a copper-deficient pasture and later fed a marginally deficient diet were studied for five months. They were divided into four equal groups; the control group received no copper supplement, groups 1 and 2 received copper as copper sulphate at 10 and 30 mg/kg of diet dry matter (DM), respectively, for five months, and group 3 received 120 mg/kg of diet dry matter for 10 days. Plasma copper concentration and the activity of erythrocyte superoxide dismutase (eSOD) were measured at the beginning of the experiment, in the cows and calves during weeks 1 and 3 after calving, and in the calves before they were turned out to pasture at a mean (sd) age of 51 (26) days. In spite of the low dietary copper content (4.2 mg/kg of DM), the plasma copper concentration of the control cows increased during the winter. All the copper supplements resulted in normal and similar plasma copper concentrations in the cows after calving, but the concentration decreased slightly between weeks 1 and 3 after calving in the group supplemented for 10 days. The treatments did not affect the eSOD of the cows. The calves born to the four groups showed the same patterns of plasma copper and eSOD. Compared with the cows, the calves had low plasma copper concentrations at week 1 and values in the normal range at week 3; their eSOD was high at weeks 1 and 3 but decreased after week 3.

Discrimination and identification of coastal Douglas-fir clones using needle flavonoid fingerprints.

This paper describes a method for discriminating and identifying 10 successful Douglas-fir (Pseudotsuga menziesii var. menziesii) clones using foliar flavonoids. All the 101 individuals analyzed by high performance liquid chromatography contained two proanthocyanidins: prodelphinidin and procyanidin and six flavonols: myricetin, quercetin, larycitrin, kaempferol, isorhamnetin and syringetin, but in different proportions. The experimental protocol used was very reproducible since the variation coefficients for each flavonoid did not exceed 9%. Submission of the flavonoid data to multivariate discriminant analysis allowed excellent discrimination of the 10 clones with 89% of the individuals being well-grouped. Then a clonal bank was established in which the fingerprint of each clone is defined by its position in the multidimensional space of the discriminant analysis. The clonal identity of several unknown individuals was determined with success by projecting their flavonoid data in a subsequent discriminant analysis.

[Occipito-posterior deliveries]. / Réflexions à propos des accouchements en occipito-sacrée

The occipito-posterior position has been thought to be a presentation leading to dystocia as a result of faulty positioning of the head. Closer observation of the facts lead one to think that this kind of position is really the result of better accommodation of the fetal head to the shapes of pelvis to which one has not become used but which are normal: this means those where the antero-posterior diameters are greater. By itself the occipito-posterior position cannot give rise to true complications. It only needs a particular mechanism. In deliveries in the occipito-posterior position progress and prognosis depend less on the presentation than on the conditions, especially the anatomical conditions, which have given rise to or made more likely the position and which go with it. It is therefore these conditions that have to be recognized each time and taken note of in order to carry out the best possible treatment.

[Cardiac transplantation. Evaluation of a consecutive series of 75 grafts]. / Transplantation cardiaque. Bilan d'une série consécutive de 75 greffes.

OBJECTIVES: The global results of various series of heart transplantation (HT) are essential to assess the life expectancy provided by this technique. Due to the increasing graft shortage, it appears essential to very strictly candidates for HT. METHODS: From March 8, 1989 to December 7, 1994, 75 orthotopic Hts were performed in 62 men and 12 women (1 case of retransplantation). The mean age was 47.46 +/- 15.02 years (range: 2.5-66 years). Four patients were younger than 10 years and 22 were older than 60 years. Our series included more cases of ischaemic heart disease (36) than dilated cardiomyopathies (33), with a history of cardiac surgery in almost one quarter (20) of patients with ischaemic heart disease. RESULTS: The immediate postoperative survival rate was 94.7% with 3 deaths attributable to refractory pulmonary hypertension associated with graft failure and one death related to postoperative tamponade. Five other patients died during the following 3 months, increasing the mean global survival to 88%. After a mean follow-up of 2.1 years (maximum 5.8 years), the actuarial 5-year survival rate was 56.8%. Eleven patients died between 4 and 38 months (mean: 18.2 months). Two deaths were due to cancers, 4 were due to septicaemia, another 4 were due to rejection and finally 1 was due to meningeal haemorrhage. The frequency (19) of reoperations for clot removal was due to the large number of patients with a history of previous heart surgery (20). CONCLUSION: Strict recipient selection, possibly based on 123I-MIBG scintigraphy, the use of pulsatile circulatory assistance systems, improved CMV, morphometry and donor-recipient age matching, should optimize the results of a technique, whose efficacy is confirmed in this series.

[Antibiotic prophylaxis in cardiac surgery: 12-hour cefazolin versus 36-hour cefazolin]. / Antibioprophylaxie en chirurgie cardiaque: céfazoline "12 h" versus céfazoline "36 h".

A prospective randomized study was carried out to assess two protocols of antibiotic prophylaxis in patients undergoing cardiac surgery with cardiopulmonary by-pass. Each patient of the first group received four intravenous injections of 1 g cefazolin over a period of 12 h, whilst in the second group each one received twelve doses over a period of 36 h. Between May 1983 and April 1984, 159 patients scheduled for cardiac surgery entered the study. Those who underwent emergency surgery, or weighed less than 20 kg, or received antibiotics in the week before surgery, or had a previous history of anaphylaxis to cephalosporins were not included in the study. Both groups were similar in their distribution of age, weight, height, sex, past history of infectious disease, surgery and iatrogenic factors. There were no significant differences between the two groups in the number of minor infections and bacterial floral changes. There were no major infections either (septicaemia, mediastinitis, endocarditis). Temperature charts were the same on the first four days. The length of stay at hospital was the same in both groups. Since the efficacy of the two protocols in preventing major infections was the same, the authors recommend the short prophylaxis with cephazolin for cardiac surgery patients.

[Contribution of autologous blood transfusion in cardiac surgery in the adult]. / Apport de l'autotransfusion sanguine en chirurgie cardiaque de l'adulte.

The use of autologous blood transfusion in cardiac surgery is still controversial. This study was prospectively designed to evaluate the haemodynamic and haematological benefits of this method, with special attention to its impact on reducing bank blood requirements. Between November 1983 and October 1984, 160 patients underwent cardiac surgery with extracorporeal circulation and were randomly assigned to two groups: group I (81 patients) was the control group and group II (79 patients) received autologous transfusion following extracorporeal circulation. Blood was withdrawn immediately after the induction of anaesthesia via a jugular catheter and stored in CPD solution at room temperature. The volume of blood removed was replaced with gelatin solutions; after bypass, blood was returned to the patient. There was no difference in systolic, diastolic or mean blood pressures between the two groups. Right atrial pressure and heart rate were not statistically different in both groups. Myocardial perfusion and myocardial oxygen consumption remained unchanged in group II compared with group I. Complete haematological evaluation was carried out before and during bypass, and thereafter daily for the first twelve days of the postoperative period. There was no significative difference between the two groups in platelet counts, fibrinogen levels, prothrombin and partial thromboplastin times. During extracorporeal circulation, mean haematocrit was 22.9 +/- 0.4% in group II and 25.3 +/- 0.5% in group I (p less than 10(-3)). The mean haematocrit time course was similar in both groups during the postoperative period and returned to preoperative value at discharge.(ABSTRACT TRUNCATED AT 250 WORDS)

[Heart surgery in 2 previously pneumonectomized patients]. / Chirurgie cardiaque chez deux patients antérieurement pneumonectomisés.

Two cases of cardiac surgery under cardiopulmonary bypass in previously pneumonectomized patients are reported. In one case a postoperative tension pneumothorax required an emergency drainage. No other complications occurred. The authors describe their technique and underline the value of an adequate pre-operative respiratory preparation and a systematic drainage of the pleural cavity.

[Complications of surgical treatment of interventricular communications in the infant]. / Complications du traitement chirurgical des communications interventriculaires du nourrisson.

Interventricular communication in the infant is presently treated by complete surgery under extracorporeal circulation and hypothermia. Improved techniques in surgery, anesthetics and resuscitation explain a low, acceptable, level of postoperative morbidity; it is estimated by most authors at 3-4% during the first six months of life. Postoperatively, acute pulmonary arterial hypertension is to be feared, resulting in low cardiocirculatory flow from right ventricle failure. In a group of 20 infants who underwent IVC surgery, the authors have observed two cases of this complication, one being lethal. Its treatment is based on the administration of alpha-blocking vasodilating agents, which can be associated or not with exogenous bêta I-adrenergic catecholamines. Results could probably be improved by preventing such acute postoperative pulmonary arterial hypertension.

[Preventive antibiotics in cardiac surgery: cefazolin versus cefamandole]. / Antibioprophylaxie en chirurgie cardiaque: céfazoline versus céfamandole.

A prospective, randomized study was carried out to evaluate two antibiotic prophylactic regimens for patients undergoing cardiac surgery with cardiopulmonary bypass. Each patient of the first group (cefazolin) received four intravenous injections of 1 g cefazolin during 12 hours, patients of second (cefamandole), four doses of 750 mg. 155 patients scheduled for cardiac operation were included in the study. (May 1983 to April 1984). Patients were not admitted to the study in case of emergency, if their weight was less than 20 kg, if they had received antibiotics during the week before surgery or if they had a history of anaphylactic reactions to cephalosporins. There were no differences between the two groups on age, weight, height, sex, previous history of infectious disease, surgery and intensive care. There were no significant differences between the two groups in minor infections. The rate of urinary tract infection by streptococci was significatively higher (p less than 0.02) in the cefamandole group (38.3%) than in the cefazolin group (17.6%). There were no major infections (septicemia, mediastinitis, endocarditis). Patients temperature was the same during the first four postoperative days. Hospital stay was the similar in the two groups. The two antibiotics are similarly effective to prevent major infections in cardiac surgery. However cefazolin was preferred for antibiotic prophylaxis in cardiac surgery because of the higher rate of streptococcal urinary infections in patients given cefamandole.

[Contribution of hemoconcentration in the recovery of residual blood from extracorporeal circulation circuits]. / Apport de l'hémoconcentration dans la récupération du sang résiduel des circuits de CEC.

The effects of recuperation from the remaining pump circuit blood with restitution to the patient after hemoconcentration are studied in 80 patients undergoing cardiopulmonary by-pass. This population is randomized into two groups: group 1 of 41 patients represents the control group and is compared with the group 2 of 39 patients who undergo the post-by-pass hemoconcentration. The volume of restored blood is 669 +/- 14 ml with an hematocrit of 38 +/- 6,6% and the total protein concentration is 81,2 +/- 2 g.l-1. The heparin level is high (4,24 units.ml-i) and the perfusion of the hemoconcentrated blood must be preceded by a slow injection of 0,8 mg.kg-1 of protamine sulfate. The advantages of this method represent a saving of two homologous blood units (p less than or equal to 10(-4] and the post operative urine output is increased by 32% (p less than or equal to 0,05). No complication, particularly infectious, was observed in this study.

[Antibiotic prophylaxis using a combination of pefloxacin and fosfomycin in heart surgery with CEC (extracorporeal circulation) in patients allergic to beta-lactams]. / Antibioprophylaxie utilisant l'association péfloxacine-fosfomycine en chirurgie cardiaque avec CEC chez les patients allergiques aux bêta-lactamines.

This study conducted for 15 months, was carried out in 34 patients with beta-lactam allergy scheduled for open heart surgery. In the study, pefloxacin was given orally an hour before the induction of anaesthesia and then as a short infusion following induction. When the bypass was stopped, pefloxacin (400 mg) and fosfomycin (60 mg.kg-1) were given in association by two separate slow intravenous infusions just before sternal closure and repeated in intensive care unit postoperatively. The antibiotic kinetics was observed in blood and cellular concentrations (atria, sternum and mediastinal part of pleura). The antibiotic level analysis showed a good diffusion during the surgical procedure, particularly during the bypass. The pefloxacin given orally was found to achieve satisfactory plasma levels of 5.4 to 6.9 mg.l-1 during sternotomy and always higher than 3 mg.l-1 during bypass. At the sternal closure, the residual plasma level was about 2.8 mg.l-1 before the reinfusion. The kinetic evaluation of fosfomycin has also shown same effective levels. Hence, the clinical potency of these antibiotics was confirmed as predicted by their excellent tissue diffusion. Thus, clinical evaluation was in favour of this antibiotic-association in most cases, except the two following ones. The first case had a lethal bronchiolar and lung reinfection with Pseudomonas aeruginosa and Candida albicans germs which appeared at the 6th postoperative day. The second patient is a case of antibiotic prophylaxis failure. He had developed an acute suppurating mediastinal infection at the seventh postoperative day with a methicillin resistant Staphylococcus aureus which had become pefloxacin fosfomycin resistant. However, the evolution was ultimately good after surgical disinfection of sternotomy and 30 days of drainage and irrigation with antiseptic solution associated with well adapted antibiotic treatment: vancomycin, pristinamycin and rifamycin. In fact, the choice of pefloxacin and fosfomycin for prophylaxis against staphylococcus in cardiac surgery is the right choice for patients having beta-lactam allergy. Their spectral activity and pharmacokinetics give us satisfactory results. But it is not the absolute solution as the bacteria responsible for nosocomial infection (hospital borne infection) may be found resistant to this association.