A simplified method of counting microorganisms in studying human autoflora

Formulae for the calculation of the count of microorganisms isolated from natural bacterial biocenoses of a child's body have been derived. The authors suggest a variant of simplified drip method for the computation of microorganism colonies in solid media. The described method helps cut down the nutrient media consumption at least 3-fold and is time-saving.     

Quality assurance and certification program for an aminoglycoside monitoring service

A Quality Assurance Program (QAP) should both evaluate and improve the quality of a service. In order to train newly employed pharmacists and ensure provision of a consistently high level of clinical service, a pharmacist training program for an Aminoglycoside Monitoring Service (AMS) and a QAP involving pharmacist certification was established. The certification program consists of a pretest, a reading/information package, an "on the job" training requirement and a posttest which pharmacists work through at their own speed. Certification requires completion of 45 hours of supervised AMS activity and a score of 90 percent on the posttest. Yearly recertification is required. As an integral part of the QAP, the clinical coordinator reviews the AMS monitoring forms monthly for specific performance standard indicators. Problems are identified and dealt with on an individual basis. The program is not mandatory, however, all pharmacists have elected to complete certification. Seven pharmacists and three pharmacy residents have participated in the certification program. All seven pharmacists and one resident received certification. A questionnaire completed by the pharmacists indicated that all felt certification was necessary and contributed to standardization and consistency of the AMS.     

Diagnosis of the pancreonecrosis using the gas chromatography method

In 62 patients with various forms of an acute pancreatitis (AP) the contents of methane, oxygen, nitrogen, carbon dioxide gas in the blood, using the gas chromatography method, was studied up. In 55 patients with pancreonecrosis the changes of concentration of methane, oxygen and nitrogen had correlated with the disease course severity, the data of ultrasound investigation and bolus computer tomography. Basing on the investigations performed in 62 patients with an AP, in 12--with other acute surgical diseases of abdominal cavity and in 20 healthy persons, it was established the possibility of determination of the methane level in the blood as a criterion of the various forms of pancreonecrosis diagnosis and the method of its diagnosis was proposed.     

Minimally invasive interventions for concurrent diseases of extrahepatic biliary ducts and pancreas

The results of treatment of 689 patients with calculous cholecystitis and of 12 with cholecystogenic acute pancreatitis (AP), to whom laparoscopic interventions were performed, are adduced. In 50 patients the indexes of operative stress were studied up. The laparoscopic cholecystectomy efficacy was established in patients with cholecystogenic AP in the absence of obturational jaundice.     

Scanning surface confocal microscopy for simultaneous topographical and fluorescence imaging: application to single virus-like particle entry into a cell

We have developed a method for simultaneous recording of high-resolution topography and cell surface fluorescence in a single scan which we call scanning surface confocal microscopy. The resolution of the system allows imaging of individual fluorescent particles in the nanometer range on fixed or live cells. We used this technique to record the interaction of single virus-like particles with the cell surface and demonstrated that single particles sink into the membrane in invaginations reminiscent of caveolae or pinocytic vesicles. This method provides a technique for elucidating the interaction of individual viruses and other nanoparticles, such as gene therapy vectors, with target cells. Furthermore, this technique should find widespread application for studying the relationship of fluorescently tagged molecules with components of the cell plasma membrane.     

Effects of the inhibitors of energy metabolism,lonidamine and levamisole,on 5-aminolevulinic-acid-induced photochemotherapy

The ability of endogenously synthesized protoporphyrin IX (PpIX) to damage Chinese hamster lung fibroblasts of the line V79 by exposure to light was examined. This treatment induced reduction of cellular ATP, GTP, of the NADH/NAD⁺ ratio and of oxygen consumption. The present results indicate a close relationship between inhibition of respiration of irradiated cells and their ability to survive, e.g. 1 min of light exposure induced 90% inhibition of oxygen consumption and inactivation of approximately 95% of the cells, while the cellular content of ATP was reduced by only 15%. This indicates that the mitochondria are one of the primary targets of 5-aminolevulinic acid (ALA)-mediated photochemotherapy (PCT). In the present study, ALA-PCT was combined with the modulators of the glycolysis and the respiration chain, levamisole (LEV) and Ionidamine (LND). A synergistic effect of combining ALA-PCT with non-toxic concentrations of LND was observed when LND was given prior to light exposure. This synergism was observed despite a substantial LND-induced inhibition of PpIX formation. At increasing doses of LND (>0.15 mM) the combination treatment becomes less efficient. This is due to the inhibition of PpIX synthesis induced by LND. A synergistic effect of ALA-PDT and LEV was found when LEV was given prior to light exposure. This was at least partly due to an LEV-stimulated effect on ALA-induced PpIX formation. However, it is not clear from the present results whether LEV may perturb energy metabolism in V79 cells since LEV alone did not reduce the energy charge or the NADH/NAD⁺ ratio. When LEV or LND were given after ALA-PCT, these 2 treatment modalities acted in an additive or slightly synergistic manner. © 1996 Wiley-Liss, Inc.     

Different modulation by histamine of IL-4 and interferon-gamma (IFN-γ) release according to the phenotype of human Th0, Th1 and Th2 clones

Histamine, an important inflammatory mediator in allergic diseases and asthma, has been reported to have modulator effects on T cells, suggesting that the bronchial microenvironment may regulate the function of resident T cells. We examined the effect of histamine on the release of the Th2-associated cytokines IL-4 and IL-5 and the Th1-associated cytokine IFN-γ by 30 CD4⁺ T cell clones from peripheral blood or bronchial biopsy of one atopic subject. Based on the IL-4/IFN-γ ratio, the clones were ascribed to the Th2 (ratio >1), Th0 (ratio 0.1 and 1) or Th1 (ratio <0.1) phenotype. Histamine inhibited IFN-γ production by Th1-like cells (P<0.02, Kruskall–Wallis), especially from bronchial biopsy, but had no effect on IL-4 release. Regarding Th0 clones, histamine inhibited IL-4 production (P<0.02) in a dose-dependent manner and slightly inhibited IFN-γ production, but had no effect on Th2-like cells. Histamine had a heterogeneous and insignificant effect on IL-5 production. The H₂-receptor antagonist ranitidine completely reversed the inhibition of IL-4 and IFN-γ production, whereas the agonist dimaprit mimicked this effect. In contrast, H₁- and H₃-receptor agonists and antagonists had no significant effect. These data demonstrate that histamine has different effects on IL-4 and IFN-γ release by T helper cells according to their phenotype via H₂-receptors. This study extends the immunomodulatory effects of histamine which may contribute to the perpetuation of airway inflammation in asthma.     

The role of size, sequence and haplotype in the stability of FRAXA and FRAXE alleles during transmission

Factors involved in the stability of trinucleotide repeats during transmission were studied in 139 families in which a full mutation, premutation or intermediate allele at either FRAXA or FRAXE was segregating. The transmission of alleles at FRAXA, FRAXE and four microsatellite loci were recorded for all individuals. Instability within the minimal and common ranges (0-40 repeats for FRAXA, 0-30 repeats for FRAXE) was extremely rare; only one example was observed, an increased in size at FRAXA from 29 to 39 repeats. Four FRAXA and three FRAXE alleles in the intermediate range (41-60) repeats for FRAXA, 31-60 for FRAXE) were unstably transmitted. Instability was more frequent for FRAXA intermediate alleles that had a tract of pure CGG greater than 37 although instability only occurred in two of 13 such transmissions: the changes observed were limited to only one or two repeats. Premutation FRAXA alleles over 100 repeats expanded to a full mutation during female transmission in 100% of cases, in agreement with other published series. There was no clear correlation between haplotype and probability of expansion of FRAXA premutations. Instability at FRAXA or FRAXE was more often observed in conjunction with a second instability at an independent locus suggesting genomic instability as a possible mechanism by which at least some FRAXA and FRAXE mutations arise.