Electrophysiologic and electrocardiographic aspects of the transplanted heart

The authors suggest that academic institutions build business-oriented policies and practices into the development of any nurse-run clinic to set the stage for financial independence when special or development funding ends. One university-affiliated program that provides 4,000 to 5,000 annual visits drastically changed its strategies when threatened with closure after free rent and other subsidies were withdrawn. The growing emphasis on ambulatory care roles for nurses at all levels makes such clinics critical to the success of the broad-based curricula of nursing education programs, as well as the clinic's value to communities they serve. Funding difficulties frequently threaten the existence of such nurse-run clinics once the initial grant funding is no longer available. This has caused a new emphasis on running such clinics in a business-wise manner. Among the strategies initiated were: direct full-pay at the time of service; a realistic business management plan; aggressive planned marketing; contracts and agreements with other agencies; obtaining provider status with selected HMOs.     

Minimal inhibition concentrations of selected antibiotics for strains of Staphylococcus aureus

Minimal inhibition concentrations (MIC) of gentamycin (Ge), neomycin (Neo), rifampicin (Rif), ampicillin (Amp), lincomycin (Lin), erythromycin (Ery), and streptomycin (STM) were determined by the agar dilution technique using 46, 130, 131, 125, 140, 139 and 142 strains of Staphylococcus aureus, respectively. The strains, selected from the collection of the authors' laboratory, were isolated from mammary gland secretions of cows affected with clinical or subclinical mastitis. The following ranges of MIC (micrograms/ml) were assessed for the antibiotics under study: Ge 0.125-0.50, Neo 0.06-0.50, Rif 0.0039-0.030, Amp 0.015-1.00, Lin 0.25-1.00, Ery 0.06-0.25, STM 0.50-64.0. Modal MIC (micrograms/ml) were as follows; Ery 0.125 (86%), Lin 0.5 (71.4%), Rif 0.007 (68.7%), Ge 0.25 (56.5%), STM 1.00 (54.2%), Neo 0.25 (53.8%), Amp 0.06 (41.6%). The order of efficiency expressed in MIC 90 (micrograms/ml) was as follows: Rif (0.015), Ery (0.125), Ge (0.25), Neo (0.25), Amp (0.5), STM (4.0).     

Detection and identification of mycobacteria in formalin-fixed, paraffin-embedded tissues by nested PCR and restriction enzyme analysis

A novel assay based on a nested PCR and restriction enzyme analysis of the PCR products was developed for the rapid detection and identification of Mycobacterium bovis and M. avium-M. intracellulare species in formalin-fixed, paraffin-embedded tissue (PET) specimens. On the basis of the nucleotide sequence data obtained in the present study, general nested primers were constructed to amplify a 424-bp segment of the gene encoding the 65-kDa surface antigen of mycobacteria. The nested PCR assay proved to be highly sensitive, since as little as 5 to 10 fg of extracted mycobacterial DNA was detected. The safety of the assay as a routine method for the diagnosis of M. bovis and M. avium-M. intracellulare in PET specimens was provided by taking various precautions. In order to prevent false positivity, specific tools and procedures were applied. To detect false-negative results and assess the efficiency of the PCR, an internal standard molecule of amplification was constructed. The digestion of the amplicons with the restriction endonuclease Sau96-I allowed the identification of M. bovis and M. avium-M. intracellulare in a large number of clinical specimens. The present results indicate that PCR combined with an internal control of amplification and restriction enzyme analysis of the amplicons provides a rapid, sensitive, and reliable method for routine diagnostic laboratories to detect and identify M. bovis and M. avium-M. intracellulare in PET specimens.     

Sources of GABAergic input to the inferior colliculus of the rat

We have studied the GABAergic projections to the inferior colliculus (IC) of the rat by combining the retrograde transport of horseradish peroxidase (HRP) and immunohistochemistry for gamma-amino butyric acid (GABA). Medium-sized (0.06-0.14 microliter) HRP injections were made in the ventral part of the central nucleus (CNIC), in the dorsal part of the CNIC, in the dorsal cortex (DCIC), and in the external cortex (ECIC) of the IC. Single HRP-labeled and double (HRP-GABA)-labeled neurons were systematically counted in all brainstem auditory nuclei. Our results revealed that the IC receives GABAergic afferent connections from ipsi- and contralateral brainstem auditory nuclei. Most of the contralateral GABAergic input originates in the IC and the dorsal nucleus of the lateral lemniscus (DNLL). The dorsal region of the IC (DCIC and dorsal part of the CNIC) receives connections mostly from its homonimous contralateral region, and the ventral region from the contralateral DNLL. The commissural GABAergic projections originate in a morphologically heterogeneous neuronal population that includes small to medium-sized round and fusiform neurons as well as large and giant neurons. Quantitatively, the ipsilateral ventral nucleus of the lateral lemniscus is the most important source of GABAergic input to the CNIC. In the superior olivary complex, a smaller number of neurons, which lie mainly in the periolivary nuclei, display double labeling. In the contralateral cochlear nuclei, only a few of the retrogradely labeled neurons were GABA immunoreactive. These findings give us more information about the role of GABA in the auditory system, indicating that inhibitory inputs from different ipsi- and contralateral, mono- and binaural auditory brainstem centers converge in the IC.     

Detection of antibiotic resistance in Enterococcus sp. Comparison of GPS-TA (BioMérieux-Vitek), Uniscept MIC-3 (bioMérieux-Vitek) and conventional methods

BACKGROUND: The treatment of severe enterococcus infections requires synergism of a beta-lactamic or glycopeptide and a aminoglycoside, but when resistance to first one or high-level resistance to aminoglycosides are present, synergism would be lost. We compared the adequacy of two commercially available systems to detect antibiotic resistance. METHODS: We studied 158 isolates of Enterococcus sp., with high-level resistance to gentamicin (40 isolates) and streptomycin (89 isolates), resistance to ciprofloxacin (34 isolates), resistance to ampicillin (7 isolates) and with intermediate susceptibility to vancomycin (3 isolates). No one was beta-lactamase producer by Cefinase disk method. We use disk diffusion as reference technique to detect high-level streptomycin resistance. The susceptibility to the remainder antibiotics was studied by agar dilution method, according to NCCLS. We studied the accuracy of GPS-TA cards and Uniscept MIC-3 in relation to the degree of agreement with conventional means, following FDA criteria. RESULTS: Essential agreement for MIC was less than 90 with MIC-3 for ampicillin (81.5%) and ciprofloxacin (71.3%). Categorical agreement rate was less than 90% (76.4%) and major error rate was higher than 3% (10.9%) with the use of MIC-3 for ciprofloxacin. Very major errors for ampicillin, vancomycin and ciprofloxacin were not produced by any system. The very major error rates for high level resistance to gentamicin and streptomycin with GPS-TA card were 5 and 15.7%, respectively. CONCLUSIONS: We do not recommend the use of the Uniscept MIC-3 panel with visual reading to detect susceptibility to ciprofloxacin. Detection of high levels of aminoglucoside resistance by GPS-TA card should be supplemented with conventional techniques because of the high rate of major error. Due to the low number of strains that have been studied, we can not assure the suitability of these systems to detect ampicillin or vancomycin resistance.     

Liver fibrosis

Surfactant abnormalities may contribute to the impairment of gas exchange observed in Pneumocystis carinii pneumonia. Analysis of rat bronchoalveolar lavage (BAL) lipid extracts from normal controls, steroid controls, trimethaprim-sulfamethoxazole (TMP-SMX) controls, TMP-SMX/P. carinii pneumonia controls, and P. carinii pneumonia animals reveal similar total phospholipid and total protein levels. However, there was a marked reduction in phosphatidylglycerol (PG) from the BAL of P. carinii pneumonia rats as compared with control animals, with a decrease from 4.91 +/- 1.29 nmol/mg protein to 0.46 +/- 0.57 nmol/mg protein (p<0.05) and a decrease, as a percent of total phospholipids, from 7.7% +/- 0.88% to 0.91% +/- 0.59% (p<0.001). Furthermore, in vitro surface activities of BAL lipid extracts from control and P. carinii pneumonia rats revealed minimum surface tension increases from 9.38 +/- 1.71 mN/m in controls to 16.36 +/- 0.83 mN/m in P. carinii pneumonia rats (p<0.05) and likewise maximum surface tension increases from 22.14 +/- 4.34 mN/m to 38.57 +/- 2.07 mN/m (p<0.01). Of interest, the surface activity of PG-deficient P. carinii pneumonia BAL lipid extracts is completely restored to that of normal controls by the addition of exogenous PG. These findings suggest that a functionally abnormal surfactant occurs in P. carinii pneumonia and that this may account, in part, for the impairment of gas exchange observed in this disorder.