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PubMed1977

Bacteriostatic and bactericidal activities of selected beta-lactam antibiotics studied on agar plates.

A novel and time-saving method for assessing bactericidal activities of beta-lactam antibiotics on agar plates is described. Minimal inhibitory concentrations (MICs) were determined by the agar dilution method. A potent beta-lactamase solution was sprayed onto the plates to inactivate the antibiotic. After further incubation at 37 degrees C overnight, the minimal concentration at which no visible growth occurred on the plates was defined as minimal bactericidal concentration (MBC). With undiluted culture as the inoculum, strains of Staphylococcus aureus, Escherichia coli, and Klebsiella pneumoniae showed a marked increase in MBC values compared with the values of MIC. There was a marked decrease in both the MICs and MBCs with diminution of inoculum size. The two concentrations were almost the same when the inoculum size was decreased to a 10(-4) dilution. In contrast, MIC and MBC for enterococci showed no marked decrease with decrease in inoculum size. Although the present study was performed with beta-lactamase-unstable penicillins and cephalosporins, the method can be applicable with any beta-lactam antibiotic if optimal and potent enzymes are available.

Ampicillin↗
PubMed2006

The signal-averaged electrocardiogram of atrial activation in patients with or without paroxysmal atrial fibrillation.

OBJECTIVE: To analyze the parameters of the time domain P-wave signal-averaged electrocardiogram (P-SAECG) and compare them with the P-wave duration on the conventional electrocardiogram (P on ECG) as well as the left atrium diameter (LAD) and left ventricular ejection fraction (EF) obtained on the echocardiogram in order to evaluate patients with paroxysmal atrial fibrillation (PAF). METHODS: One hundred and eighty-one patients were included in the study: 117 with confirmed PAF and 64 without PAF. The P-SAECG parameters used were: the filtered P-wave duration (FPD), the root mean square (RMS) voltages in the last 40, 30 and 20 ms of the filtered P-wave (RMS 40, RMS 30 and RMS 20), the root mean square voltage of the filtered P-wave potentials (RMS P), the integral of the potentials during the filtered P-wave (Integral P) and the filtered P-wave late potential durations below 3 microV (PL<3). RESULTS: The parameters that presented significant statistical differences between the groups were: FPD, RMS 40, 30 and 20, PL<3, P on ECG and LAD. The ROC curve calculations demonstrated the best cut-off points and performance estimates for each parameter: sensitivity, specificity, area under the curve and p-value (p). CONCLUSION: The time domain P-SAECG proved to be a superior method to identify patients with paroxysmal atrial fibrillation than the conventional electrocardiogram and echocardiogram.

Adolescent↗
PubMed1983

[Detection of learning impairment in offspring in reproduction tests].

We examined the ability of learning and its retention employing test with the water T-maze, shuttle box and passive avoidance response (PAR) in offspring from mother rats treated with N-methylnitrosourea (MNU) or chlorpromazine (CPZ). The offspring of MNU-induced learning impairments were detected only in tests with the water T-maze and PAR. On the other hand, CPZ induced impairments were detectable only in tests with the shuttle box and PAR. Thus offspring from mother rats treated with MNU or CPZ showed partially different responses. In retention tests, both MNU and CPZ caused impaired ability in the water T-maze and PAR. In the PAR test, impairment was most clearly detectable. From these results, the PAR test appears to be the most useful method in detection of impairments in learning and retention ability, especially the latter, in the reproduction test of offspring.

Animals↗
PubMed1984

Prophylactic effect of neuroleptics in symptom-free schizophrenics: a comparative dose-response study of haloperidol and propericiazine.

Remitted schizophrenic outpatients were treated in order to prevent relapse with three doses of haloperidol or propericiazine for 1 year in a double-blind controlled study employing a randomized design. The drug's ability to prevent relapse was evaluated by counting the number of symptom-free days for each patient before any sign of relapse or over-dose appeared. Patients were randomly assigned to the following drugs orally administered once per day at night: placebo; haloperidol 1 mg, 3 mg, and 6 mg; propericiazine 10 mg, 30 mg, and 60 mg. Serum prolactin levels in each patient were estimated by radioimmunoassay. All patients treated with placebo relapsed within 1 year and the relapse rate with placebo was significantly higher than with any dose of the two neuroleptics. Haloperidol increased the number of symptom-free days in a dose-dependent manner. Propericiazine at 10 mg and 30 mg also increased the number of symptom-free days dose-dependently but at 60 mg, the number decreased. It appears that propericiazine shows an inverted U-shaped dose-response curve. Prolactin levels were elevated dose-dependently by both drugs but failed to show a significant correlation with the number of symptom-free days. The present results indicate that haloperidol is superior to propericiazine from the viewpoint of the wider "therapeutic window" in maintenance treatment and antidopaminergic properties of neuroleptics, wherein it is important to prevent relapse even in remitted schizophrenics.

Adult↗
PubMed1992

Efficacy and tolerability of moclobemide compared with fluvoxamine in depressive disorder (DSM III). A French/Swiss double-blind trial.

The efficacy and tolerability of moclobemide and fluvoxamine, two new types of antidepressant agents, were compared in a multicentre, double-blind prospective study of patients with a diagnosis of major depressive episode (DSM III). Patients were randomized to receive either moclobemide (150 mg) or fluvoxamine (50 mg) twice daily for 7 days, immediately following a washout period of at least 1 week. Dosages were increased where necessary on day 8, to a maximum of moclobemide 450 mg or fluvoxamine 200 mg and in most cases were maintained at these levels for the remainder of the study period (4-6 weeks). Both treatment groups showed a marked antidepressant effect. While both treatments were well tolerated, moclobemide showed a more favourable side-effect profile than fluvoxamine. Of the 126 patients eligible for evaluation, 34 withdrew from therapy, 22% in the moclobemide group and 30% in the fluvoxamine group. Adverse events were reported in 41.8% of patients treated with moclobemide compared to 60.3% of patients in the fluvoxamine group. Reports of dry mouth and other anticholinergic effects were more frequent among those treated with fluvoxamine. A greater number of gastrointestinal complaints, especially nausea, also occurred in the fluvoxamine-treated patients.

Adult↗
PubMed2000

Early feeding after oncological colorectal surgery in Japanese patients.

We aimed to determine the safety and feasibility of an alternative postoperative oral intake protocol for Japanese patients who had had oncological colorectal surgery, with the goal of shortening the length of postoperative stay to the same level as that in Western countries. The subjects of the study were 42 consecutive patients who underwent elective oncological colorectal surgery. Two protocols., Japanese traditional management (TM: n = 20) and alternative management (AM; n = 22), were employed. The two protocols differed in their management of oral intake after surgery. Tolerance of the alternative protocol and the incidence of postoperative complications were determined. The mean length of postoperative stay in the AM and TM groups was 11.5 +/- 1.2 and 24.0 +/- 2.1 days, respectively (P < 0.0001; t-test). Analysis of the patients who underwent open operation also revealed that the length of postoperative stay was less in the AM group than in the TM group (12.3 +/- 1.7 and 24.7 +/- 2.2 days, respectively, P = 0.0003). One patient in the TM group developed small bowel obstruction during the introduction of oral intake. One patient in the AM group also developed small bowel obstruction, after discharge. and was readmitted. Other patients tolerated both the TM and AM protocols. Morbidity in the AM group was 20.0%, and that in the TM group was 30.0% (P = 0.72). The employment of an alternative postoperative oral intake protocol was safe and feasible for Japanese patients. Further investigation is needed to determine the possible benefits, both medical and socioeconomic, of this approach.

Adult↗
PubMed1989

Microsurgical reversal of female sterilization. Long-term follow-up of 117 cases.

A prospective study of 117 consecutive microsurgical reversals of female sterilization from April 1981 to December 1984 was carried out to determine which factors affected the pregnancy outcome. Nylon sutures, 8-0, were placed through the muscularis and mucosal layers. A short abdominal incision was made. The patients were followed for 3.5 to 7.0 years. The term delivery, intra-uterine pregnancy, spontaneous abortion and ectopic pregnancy rates were 81.2%, 83.8%, 1.7% and 1.7%, respectively; two ectopic pregnancies occurred 14 and 24 months after the reversal procedures. We found that the time interval between sterilization and reversal and the methods of sterilization affected the pregnancy outcome. The success rate with intervals of less than five years was much higher than with intervals of more than five years; Pomeroy cases were more reversible than Uchida cases. We did not observe any influence of the anastomosis site on the pregnancy results. The term delivery rates for isthmus-isthmus, isthmus-ampulla and ampulla-ampulla anastomoses were 78.8%, 80.0% and 84.6%, respectively. Patients should be followed for two years at least to determine the pregnancy outcome as well as the occurrence of ectopic pregnancy.

Anastomosis, Surgical↗
PubMed1992

Recombination and amplification of multiple portions of genomic DNA by a modified polymerase chain reaction.

A novel method, a modified polymerase chain reaction (mPCR), is reported for in vitro recombination and amplification of two or more regions of DNA located in distinct parts of a genome such as exons of a gene. The reaction consists of three stages. In the first stage, the portions are amplified individually with the aid of four or more primers. Of the primers, two (outside primers) are complementary to 3'-terminal parts of the objective, recombinant DNA, and the others (inside primers) are complementary to 3'-terminal parts of the other portions. Some of the inside primers have extra nucleotide sequences at their 5'-termini which are complementary to 5'-terminal parts of the portions to be recombined. In the second stage, the strands with complementary sequences in their 3'-terminal parts are annealed sequentially to one another and are converted to recombinant DNA with DNA polymerase. In the final stage, the objective DNA is amplified with the excess outside primers. The application of this method for in vitro recombination and amplification of three exons of the gene for human muscle-specific phosphoglycerate mutase is described.

Amino Acid Sequence↗
PubMed1984

Monoclonal antibodies against diphtheria toxin fragment A. Characterization and introduction into living cells.

Monoclonal antibodies against fragment A of diphtheria toxin were isolated and characterized. Three antibodies with similar affinities for fragment A had different effects on the NAD: EF2-ADP ribose transferase activity of fragment A; i.e., antibody DA1 almost completely inhibited the enzymic activity at a molar ratio of one, whereas DA2 inhibited only partially and DA3 had no effect. However, when fragment A176 from the mutant toxin CRM176 (about 1/10 as active as wild type) was used, DA2 proved a more effective inhibitor than DA1. The affinities of these antibodies for the enzymically inactive mutant fragments, A197 and A228, were significantly less manifest than for wild-type fragment A. Binding of the antibodies to whole toxin and the chain termination mutant CRM45 was weak. When DA2 was introduced into Vero cells growing in monolayers, by using the red cell ghost fusion method, the cells became resistant to CRM176. The anti-fragment A antibodies may serve as the basis of a simple method for selection of cells into which other molecules have been co-introduced.

Adenosine Diphosphate Ribose↗
PubMed1995

High-performance liquid chromatographic determination of primary aromatic amines in urine after derivatization to an azo dye with 2-aminoanthracene.

A sensitive HPLC method for the determination of primary aromatic amines (anilino compounds) is described. Samples were prepared by derivatization of the substrate to an azo dye with 2-aminoanthracene (2-AA). 2-AA was found to react with the diazonium salts prepared from substituted anilines such as 4-halo, -sulfonyl, -carboxyl, -nitro or -acetyl derivatives, but not 4-hydroxy or -alkyl derivatives. In this work, three model compounds [sulfanilamide, 4-aminobenzoyl-beta-alanine and 4-aminobenzoic acid (PABA)] were used to test the linearity and accuracy of the method. Chromatographic separation was carried out using a reversed-phase column (ODS) and ultraviolet detection at 279 nm. Good linearity for the three compounds was found within the range 50-2000 ng/ml. The intra-day coefficient of variation for the three compounds (at 100, 500, 1000 ng/ml) was below 10%. Using this method, the urinary excretion of PABA and its metabolites was studied after oral administration of PABA to rats.

Aniline Compounds↗
PubMed1997

Improved technique of heterotopic cervical heart transplantation in mice.

BACKGROUND: The method for mouse vascularized heart transplantation have been described using suture and cuff techniques. Technical problems have limited its widespread use. Here, we describe our method of modified cervical heterotopic transplantation with the cuff technique. METHODS: By using a smaller Teflon cuff (external diameter 0.6 mm, internal diameter 0.4 mm) and superfine-tip forceps, it became possible to directly pull the edge of the carotid artery and evert the proximal end of the artery over the cuff. Similarly, the external jugular vein could be easily everted over a 22-gauge cuff with this direct pulling method. RESULTS: By these modifications, the operation time was reduced. It usually takes 20 min for the donor harvest, 15 min for preparation of the cervical vessels, and 15 min for anastomosis. All procedures from the donor harvest through skin closure of the recipient mice can be completed within 1 hr, and ischemic time is within 25-40 min. CONCLUSIONS: This method can be used to investigate cyclophosphamide-induced tolerance and mechanisms of reperfusion injury.

Anastomosis, Surgical↗
PubMed1991

Identification of pre- and postcentral gyri on CT and MR images on the basis of the medullary pattern of cerebral white matter.

The authors illustrate a new method to identify the pre- and postcentral gyri on computed tomographic (CT) and magnetic resonance (MR) images of the brain on the basis of the pattern of the medullary branches of the cerebral white matter. The most commonly used method to identify the gyri depends on recognition of the central sulcus by surface arrangement of the sulci. The two methods were compared by analysis of CT images of 104 subjects who had normal findings (age range, newborn to 60 years; 57 males and 47 females). The usefulness of the new method was also determined in angiographic studies of nine patients with space-occupying lesions. The method is especially helpful for identification of gyri on the lower level of the centrum semiovale and if space-occupying lesions are present that may result in a blurred depiction of sulci. Since MR imaging depicts the medullary branches more clearly than does CT, this new method should facilitate identification of the gyri with either modality.

Adolescent↗
PubMed1993

A novel colorimetric method for determination of glycated protein based on 2-keto-glucose release with hydrazine.

We tried to measure glycated proteins by a novel method based on colorimetry of 2-keto-glucose which is released from the glycated protein (ketoamine) on heating with hydrazine. Reaction conditions were optimized with glycated human serum albumin (glc HSA) as a model compound. Ketoamine reacted quantitatively with hydrazine on heating at 100 degrees C for 0.5 h, followed by heating with phenylhydrazine at 60 degrees C for 1 h. Glucose interference with the assay was eliminated by preincubation of the sample with glucose oxidase at 37 degrees C for 0.5 h. Time courses for the coloration of glc HSA and human serum showed a profile similar to that of N-p-tolyl-D-isoglucosamine under optimized reaction conditions. The lower limit for the assay of glc HSA was 0.7 microM. The serum level of glycated proteins measured by the present method correlated well with that (fructoamine value, microM) measured by the conventional method (nitroblue tetrazolium-reducing method) (r = 0.92, n = 35). In conclusion, the present method is a novel, highly sensitive and reliable one for measuring glycated proteins in biological samples.

Colorimetry↗
PubMed1995

Optimization of preparative conditions for polylactide (PLA) microspheres containing ovalbumin.

Polylactide (PLA) microspheres containing ovalbumin (OVA) as a model protein were prepared by a water-in-oil-in-water (w/o/w) emulsion solvent evaporation method. The optimization of preparative parameters for the PLA microspheres containing OVA were performed, and the in vitro characteristics of the obtained microspheres were examined. Firstly, a smaller internal aqueous phase volume was found to be advantageous in obtaining high loading efficiency. Secondly, the addition of 2-10% (w/v) NaCl into the external aqueous phase (0.5% (w/v) polyvinyl alcohol solution) also improved OVA loading efficiency. Prepared products showed a sharp release of OVA at the initial phase, but the following phase was characterized by a slow release rate of OVA that continued at least 28 d. The release rate of OVA from microspheres made of PLA with a molecular weight of 15400 was faster than that from microspheres made of PLA with a molecular weight of 58300. However, the LA/GA (lactide/glycolide) ratio was not likely to have much effect on the release profile of OVA. Finally, the effect of PLA microsphere particle size on the release profiles of OVA was examined. The extent of burst release at the initial phase increased as the mean diameter of prepared PLA microspheres decreased. For example, the PLA microspheres with a small mean diameter (5.0 microns) showed a 40% burst release, but almost 30% of OVA remained in the PLA microspheres (confirmed by HPLC method) after the 28 d release test, suggesting the possibility of using this carrier as a long-acting protein delivery system.

Biocompatible Materials↗