Biomedical subjects
Publications and source records attributed to M García.
Colonies of Streptomyces antibioticus were studied by transmission and scanning electron microscopy. The micrographs show that substrate mycelium growth takes place among an intercellular material and this mycelium is covered by a surface film. This structure could be a boundary between the aerial mycelium and the substrate mycelium.
Human umbilical vein endothelial cells (HUVEC) were treated with recombinant interleukin (IL)-1 beta, and the metabolism of exogenous linoleic acid was studied. High performance liquid chromatography, gas chromatography-mass spectrometry, and chiral analysis revealed that HUVEC enzymatically convert linoleic acid mainly into 13-(S)hydroxy-9(Z),11(E)-octadecadienoic (13-HODE) and 9-(R)hydroxy-10(E),12(Z)-octadecadienoic acids, which may isomerize toward all-trans compounds. IL-1 beta increased the formation of all octadecanoids in a time- and dose-dependent manner with similar EC50 (approximately 1 unit/ml). The apparent Km values of linoleic acid were 15.59 +/- 8.39 and 152.9 +/- 84 microM (p < 0.05) in IL-1 beta-treated cells and controls, respectively, indicating a higher substrate affinity in cells stimulated with IL-1 beta. Ratios of S/R enantiomers for the hydroxyoctadecanoids produced by untreated and IL-1 beta-treated cells were similar to those from isolated cyclooxygenases (COXs), whereas isolated 15-lipoxygenase yielded 13-HODE with a strict S configuration. The formation of octadecanoids was inhibited in a dose-dependent manner by several COX inhibitors in both controls and IL-1 beta-treated cells, COX2 selective inhibitors being more effective on IL-1 beta-treated cells than on controls. COX1 and COX2 protein levels increased less than 2-fold and 8-fold, respectively, after IL-1 beta treatment. The specificity of COX inhibitors was proven since they did not inhibit 13-HODE formation by human polymorphonuclear leukocytes. Overall, these results indicate that COXs are responsible for the oxidative metabolism of linoleic acid in HUVEC, and IL-1 beta increases it by inducing the expression of new enzyme, mainly COX2.
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The localization of protein kinase C (PKC) was studied immunocytochemically in the Müller cells of the carp retina. Electron microscope immunocytochemistry (using a monoclonal antibody to the alpha and beta isoenzymes of PKC) showed PKC-immunoreactivity mainly inside some mitochondria, especially along the mitochondrial cristae whereas other mitochondria in the same Müller cells showed no staining. Despite a detailed analysis we did not find any significant morphological difference between labeled and unlabeled mitochondria. These results demonstrate, for the first time, the presence of PKC immunoreactivity inside mitochondria and suggest that individual mitochondria may differ in signal transduction pathway.
OBJECTIVE: To determine the long term cardiovascular effects in adolescents exposed to ritodrine during prenatal stage. STUDY DESIGN: Twenty-seven ritodrine-exposed subjects and 19 controls, of both sexes, clinically healthy, and born at normal term, were evaluated for systolic blood pressure (SBP), diastolic blood pressure (DBP) and heart rate (HR) each during 48 hours, with a Colin Ambulatory monitor. Data analysis were performed by individual Cosinor method, and non-parametric Mann Whitney's test and Lamotte procedures. RESULTS: Adolescents who were exposed to ritodrine showed losses in the HR rhythm and rising middle pulse values. Although SBP and DBP rhythm remains constant, the increase of maximum and minimum values for SBP as well as a rise of maximal rates of DBP suggest a possible hypertensive effect mediated by beta mimetic exposure. CONCLUSION: This study suggests a hypertensive effect mediated by beta mimetic exposure.
During waking h, the existence of ultradian rhythms in gross motor activity has been described in nonprimates, nonhuman primates and newborn humans, but not in adult humans. Some of the previous studies suggested that the appearance of these rhythms could be favored by conditions of isolation and low environmental demands. To confirm the existence of ultradian rhythmicity in the gross motor activity of adult humans and to describe their characteristics, an actimeter was used to record the mobility of 13 adults who remained alone and isolated for 5 h (15:00-20:00) in a monotonous environment with nothing to do. Least squares rhythmometry analysis showed that the gross motor activity of 12 out of 13 subjects had significant rhythms (p < 0.05) within the ultradian band, with periods of between 0.5 and 2.5 h. There were important individual differences between the dominant periods and the same subject might show more than one significant period. These rhythms were stable and they tended to appear immediately the experiment began. The results suggest that a philogenetically old mechanism that organizes gross motor activity in ultradian rhythms exists in adult humans.
In this paper we have applied the matrix method to calculating spectacle magnification (SM) and relative spectacle magnification (RSM) for myopic eyes corrected with ophthalmic lenses (OLs). We have been able to obtain very simple expressions of the magnifications and analogous values to the ones obtained using other geometric methods. Through the paraxial 2 x 2 object-image matrix for spherical systems, we can calculate the correcting lens pupil matrix (ML) and the corrected ametropic eye pupil matrix (Mca). These two matrices, that is ML and Mca, directly provide the lateral magnification and the lens-eye system's image focal parameters that enable us to easily calculate SM and RSM. The magnifications have been calculated for the case of myopic eyes corrected with OLs, which shows the validity of this method since its results are similar to the ones obtained using other methods. The analysis of other correcting systems such as contact intraocular lenses will be presented in two associated papers (to be published as parts II and III of this review).
BACKGROUND: Analysis of the in vitro sensitivity of clinical isolates of different yeast species to amphotericine B and two inhibitors of cell wall beta-glucane synthesis: the lipopeptide cilofungin and the liposaccharide papulacandin B, as well as the possible interactions among these antifungal types. METHODS: The in vitro sensibility of C. albicans strains and other yeast species of clinical origin was studied, using the macrodilution method in RPMI broth as recommended by NCCLS. In six C. albicans and two T. glabrata strains the interaction between amphotericine B and one of the beta-glucan inhibitors was also assessed, by combining one antifungal agent in a two-fold dilution series with subinhibitory concentrations (1/4MIC) of the second antifungal. RESULTS AND CONCLUSIONS: Of all three antifungals, amphothericine B proved to be the more active one, with a MIC90 of 1.25 micrograms/ml for C. albicans. MIC90 of cilofungin and papulacandine B is 5 micrograms/ml for both antifungals. The spectrum of action for the different yeast species is broader for papulacandin B than for cilofungin. On the other hand, no synergism has been observed between the two glucane inhibitors and amphotericine B.
BACKGROUND: Sensitivity and specificity of latex agglutination test on samples obtained by transthoracic needle aspiration was evaluated for diagnosis of pneumococcal pneumonia by using the polymerase chain reaction as reference method. METHODS: Samples from 29 patients with community acquired pneumonia were processed for culture, antigen detection and polymerase chain reaction. The latex agglutination test was performed with a reagents kit (Slidex méningite kit, BioMérieux, France) using the procedure recommended by the manufacturer. The polymerase chain reaction was carried out by using two primers: Pn2x up and Pn2x down that amplify the PBP2x gene of Streptococcus pneumoniae. RESULTS: Culture was positive in 5 patients, antigen detection in 15 and polymerase chain reaction in 14 patients. The specificity of latex agglutination test was 93% compared with polymerase chain reaction as reference method. CONCLUSION: The pneumococcal antigen detection by latex agglutination test is as sensitive as the polymerase chain reaction, it seems to be highly specific, and it is rapid and easy to perform.
The disulfide bridges of toxin 2 from the venom of the scorpion Centruroides noxius Hoffmann were found by amino acid sequence determination of fragments of native toxin, produced by enzymatic cleavage and separated by high-performance liquid chromatography (HPLC). They are: Cys12-Cys65, Cys16-Cys41, Cys25-Cys46 and Cys29-Cys48. The coordinates of the X-ray diffraction structure of toxin variant 3 of C. sculpturatus [(1980) Proc. Natl. Acad. Sci. USA 77, 6496-6500] were used to construct a three-dimensional model of toxin 2. All the amino acid replacements were easily accommodated, and the modeled structure reveals a clustered pattern of sequence variation, which may help to identify residues responsible for functional differences among toxins of mammals and insects.
In the three years between November 1989 and October 1992, an outbreak of methicillin-resistant Staphylococcus aureus (MRSA) affected 990 patients at a university hospital. The distribution of patients with carriage, colonization or infection was investigated prospectively. Nosocomial acquisition was confirmed in at least 928 patients, 525 of whom were identified from clinical specimens as being infected (n = 418) or colonized (n = 107) by MRSA. An additional 403 patients were identified from screening specimens, of whom 58 subsequently became infected and 18 colonized. Screening of the nose, throat and perineum detected 98% of all carriers. Of the 580 infections in 476 patients, surgical wound, urinary tract and skin infections accounted for 58% of the infections. Of the 476 infected patients, death was attributable to MRSA infection in 13%. Colonization with MRSA was found in 127 patients and 42% of 165 colonized sites were the skin. Auto-infection from nasal carriage or cross-infection, probably via staff hands, seemed to be the most common mode of acquisition of MRSA infections.
The in vitro expression of surface epitopes for different strains of Mycoplasma gallisepticum (MG) was studied with a panel of monoclonal antibodies (mAbs) using indirect colony immunostaining and Western blot (WB) analyses. Immunostaining of colonies with mAbs showed that five epitopes had different degrees of variable expression, while one epitope was permanently expressed in vitro. Colonies that failed to express the studied epitopes had the potential of phenotypically switching the expression of these epitopes in vitro. Variable and permanently expressed epitopes were associated with more than one protein and not all mAb-defined proteins were responsible for the immunostaining of intact MG colonies. The ability of MG to variably express their surface epitopes maybe the mechanism utilized by the microorganism to avoid the host immune response.
Policosanol is a new chemical entity composed of 8 higher aliphatic alcohols obtained from sugar cane (Saccharum officinarum), L. wax, whose cholesterol-lowering effects have been demonstrated in experimental models, healthy volunteers and patients with type II hypercholesterolemia. This study investigated the oral toxicity of policosanol administered for 52 weeks to beagle dogs. Twenty-four beagle dogs (12 males and 12 females) were distributed randomly in 3 experimental groups (4 animals/group): a control and 2 treated groups receiving policosanol at 30 and 180 mg/kg daily (7 days/week) by gavage. No mortality was observed in any group. Overall, policosanol was well tolerated throughout the study and no toxic symptoms were observed. All groups showed similar weight gain and food consumption. Lipid profile determinations showed that policosanol decreased total cholesterol by 20% approximately from 8 to 52 weeks. Cholesterol-lowering effects did not wear off during the study, thus demonstrating the persistence of the effectiveness. Triglycerides and high density lipoprotein-cholesterol (HDL-C) were not changed significantly. No blood biochemistry or histopathological disturbances attributable to treatment were observed. This study has shown that no drug-related toxicity was induced by policosanol administered up to 180 mg/kg/day for 52 weeks to beagle dogs. Since this dose is approximately 620 times higher than the maximal recommended therapeutic dose (20 mg/day) it indicates a good safety margin of this product.
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Dialysis-related amyloidosis is a type of amyloidosis which had beta 2-microglobulin as the major protein constituent and occurs predominantly in haemodialysis patients. Its prevalence is very high with increasing time on dialysis treatment and its pathogenesis is not completely understood. While remarkable progress has been made in the identification of the components of the deposits, there are no reports characterizing the cells surrounding the amyloid fibrils. To characterize the cellular composition of the amyloid material, specimens from seven patients treated by maintenance haemodialysis were studied with immunoperoxidase labelling using monoclonal antibodies to leukocytes (CD3, CD14, CD68, CD4, CD8, CD45). The results were very reproducible for the seven deposits assessed: Of the 182 +/- 26 leukocytes/0.2 mm2 of amyloid tissue expressing the 71.5-CD45 marker (common leukocyte), 91 +/- 6% were CD68 (KP1) positive (monocyte macrophage). No CD3-positive cells (T-cell marker) were found in six of the seven patients, with only 1.6% in the remaining one. The present study shows that although amyloidosis has classically been considered as an acellular pathology, clearly there are cells surrounding amyloid fibrils. Strikingly, these cells are almost exclusively macrophages; there are no lymphocytes or granulocytes. The putative role of macrophages in the pathogenesis of beta 2-microglobulin amyloidosis remains to be established. However, the identification and quantitation of the cells surrounding the amyloid deposits may be important for subsequent studies to elucidate amyloid pathogenesis and particularly protein-cell interactions.
The pattern of growth of aerial mycelium in Streptomyces species was investigated by autoradiography. Colonies of Streptomyces antibiotics were labeled with N-acetyl-D-[1-3H] glucosamine to localize the sites of hyphal growth during the development of aerial mycelium. Autoradiographs obtained with sections of the colonies revealed that hyphal growth occurs not only at the top of the colony but also in the inner zones of the aerial mycelium.
STUDY OBJECTIVE: To evaluate the safety and efficacy of transthoracic aspiration with an ultrathin needle in the microbiologic diagnosis of pulmonary infections in HIV-infected patients. DESIGN: Retrospective review of cases. SETTING: A 500-bed teaching hospital in Lleida, Spain. PATIENTS: Forty-five HIV-infected patients admitted between March 1989 and March 1993 with clinical and roentgenographic evidence of pulmonary infection and without contraindications for transthoracic needle aspiration (TNA). INTERVENTIONS: Forty-seven TNAs were performed in the emergency room (20) or during hospitalization (27). The TNA procedures were done without premedication and without fluoroscopic guidance. Specimens were processed using routine microbiologic and cytologic techniques; in addition, polymerase chain reaction (PCR) for Pneumocystis carinii was carried out since March 1992. Development of adverse effects was carefully evaluated. RESULTS: The TNA was effective in 29 (62 percent) out of 47 procedures. The diagnosis was obtained for 14 of 15 patients with P carinii pneumonia, 8 out of 14 patients with bacterial pneumonia, and 4 out of 12 patients with tuberculosis. Other pathogens recovered were Nocardia asteroides, Cryptococcus neoformans, Rhodococcus equi, and Mycobacterium avium. No false-positive results were obtained. Pneumothorax developed in eight (17 percent) procedures, but only one procedure resulted in a pleural drainage; the incidence of other adverse effects was low and clinically irrelevant. CONCLUSION: Our study suggests that TNA can be a useful technique in establishing the etiologic diagnosis of pulmonary infections in HIV-infected patients, with a good sensitivity, high specificity, and relatively low incidence of serious complications, with TNA appearing as a reliable alternative to more uncomfortable methods.
The earth of ornamental plants is one of the main reservoirs of Aspergillus type of fungi in hospital areas. We studied 174 ornamental interior plants from a hospital at Santiago. Samples were obtained from the soil surface and sowed in Sabouraud-glucose agar, adding streptomycin and G-penicillin. After 72 h of culture, at least one strain of Aspergillus was isolated from 140 samples (80.5%). The most frequently isolated strain was A fumigatus (129 samples), followed by A niger (75 samples). A fumigatus and A niger were the only isolated strains in 65 and 11 samples respectively. These findings confirm that ornamental plants can be important reservoirs of Aspergillus strains, a potential infectious agent for immunocompromised patients, in hospital areas.