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C Miralles

Publications and source records attributed to C Miralles.

15 recordsLinked to original sources

Regulation of iNOS expression and glutathione levels in rat liver by oxygen tension.

Molecular oxygen (O(2)) regulates the expression of a variety of genes. We hypothesized that O(2) tension may regulate iNOS expression in rat liver through the production of reactive oxygen species (ROS) and the reduction of intracellular glutathione (GSH) levels. To investigate this hypothesis, we determined the effects of hyperoxia upon iNOS induction (both at the protein and mRNA level) and the intracellular concentration of GSH in an isolated in vitro perfused rat liver preparation. To study the potential involvement of ROS in the intracellular signaling pathway linking changes in oxygen tension to gene expression, we repeated these determinations in the presence of the thiol antioxidant N-acetyl-L-cysteine (NAC). We found that 95% O(2) tension caused a significant induction of the iNOS protein and mRNA levels paralleled by a significant fall in intracellular GSH concentration. The addition of NAC (1 mM) to the perfusate during hyperoxia blocked the induction of iNOS and restored GSH levels. These results indicate that molecular O(2) regulates the expression of iNOS in rat liver at the transcriptional level, most likely through the production of ROS and the reduction of intracellular GSH levels.

Acetylcysteine↗

Clinicopathological characteristics of breast carcinomas with allelic loss in the p73 region.

p73, a new member of the p53 family, has been mapped to chromosome 1p 36, a region where loss of heterozygosity (LOH) is frequently observed in primary human tumors. Allelic loss studies involving the 1p arm in breast carcinomas offer rates ranging from 13% to 75%, depending on the genetic interval being studied. We investigated LOH in an intragenic microsatellite marker, and those centromerically flanking the p73 gene, at 1p 36, and their correlations with patient age and 10 pathologic parameters in a series of 193 breast carcinomas. The LOH analysis was performed by amplifying DNA by PCR, using the five markers of the 1p 36 region (p73P1, D1S2694, D1S214, D1S2666 and D1S450). LOH was found in at least one of these markers in 27% of tumors. When we established the comparison between tumors with and without LOH and the distribution of the 10 pathologic parameters considered, we observed statistically significant differences in association with higher histologic grade (p = 0.02), more advanced pathological stage (p = 0.02), peritumoral vessel involvement (p = 0.04) and poorly differentiated carcinomas (p = 0.01), as well as in tumors that concomitantly exhibited lymph node metastases, peritumoral vessel involvement and absence of steroid receptors (p = 0.02). These data suggest that LOH in the p73 region could be pathogenically related to breast cancer and possibly to a poor tumor prognosis.

Adult↗

Abnormal lipid peroxidation in patients with sleep apnoea.

The prevalence of cardiovascular diseases is increased in patients with the obstructive sleep apnoea syndrome (OSAS). The fall and rise of arterial oxygenation that follows each apnoea may increase lipid peroxidation and contributes to explaining this association. In the present study, the authors determined lipid peroxidation in patients with OSAS and the effect of treatment with continuous positive airway pressure (CPAP). Fourteen male patients with severe OSAS (59+/-5 apnoea x h(-1)) (+/-SEM) and 13 healthy nonsmoking, male volunteers of similar age were studied. Patients were studied at diagnosis and after treatment with CPAP for more than 1 yr (>4 h x night(-1)). A venous blood sample was obtained early in the morning after fasting all night. In patients with OSAS, a sample before and during sleep was also obtained. Low density lipoprotein (LDL) particles were isolated by sequential ultracentrifugation. Their level of oxidation was determined by the thiobarbituric acid assay (TBARs), and their susceptibility to oxidation by the lag phase measurement. Patients with OSAS showed higher TBARs (28.1+/-2.8 versus 20.0+/-1.8 nmol x malondialdehyde x mgLDL protein(-1), p=0.02) and shorter lag phase values (83.8+/-3.4 versus 99.7+/-3.4 min, p=0.005) than controls. These differences were not due to the smoking status of the patient. Likewise, these values did not change significantly throughout the night yet, the lag phase value was significantly improved by treatment with CPAP (124.9+/-8.5 min; p<0.001). These results indicate that obstructive sleep apnoea syndrome is associated with abnormal lipid peroxidation and that this is improved by chronic use of Continuous positive airway pressure. These results can contribute towards explaining the high prevalence of cardiovascular diseases seen in Obstructive sleep apnoea syndrome.

Adult↗

Changes induced by oxygen in rat liver proteins identified by high-resolution two-dimensional gel electrophoresis.

Molecular oxygen (O2) regulates the expression of a variety of genes. Several of the proteins that respond to changes in oxygen concentration have been identified in a variety of cell lines. We extend these previous studies by analyzing the effect of oxygen on the entire protein expression profile of an intact organ using high-resolution two-dimensional gel electrophoresis. To this end, we used an isolated, in vitro perfused organ preparation to produce two groups of rat livers perfused with high (95% O2, 5% CO2) or low (95% N2, 5% CO2) oxygen concentrations. Using two-dimensional gel electrophoresis we compared the protein expression profiles of both groups of livers. Computer analysis of the files obtained after laser densitometry of the two-dimensional gels revealed two spots that were strongly up-regulated in high PO2 perfused livers compared with low PO2 perfused livers. These spots were analyzed by peptide mass fingerprinting analysis. These spots were identified as arginase 1 (liver-type arginase; EC 3.5.3.1) and mitochondrial enoyl-CoA hydratase 1 (EC 4.2.1.17). The possible role of these proteins in its new context of oxygen availability is discussed.

Animals↗

Analytical and clinical evaluation of TSH and thyroid hormones by electrochemiluminescent immunoassays.

OBJECTIVES: To perform an analytical evaluation of the new electrochemiluminescent immunoassays (ECLIA) for TSH, FT4, and T3 in the Elecsys 2010 immunoassay system. To assess the clinical classification of patients under suspicion of thyroid disease based on these laboratory assays. MATERIALS AND METHODS: The analytical evaluation included the performance of minimum detectable concentrations, within-assay and inter-assay precision for the three analytes, functional sensitivity and linearity studies for TSH, and method comparison with the previous methods of RIA for FT4 and T3, and IRMA for TSH in current protocols of our institution. 102 patients with clinical suspicion of thyroid disease were assayed by ECLIA and radioactive techniques. Their differential clinical classification based on laboratory tests was studied as well. RESULTS: The minimum detectable concentrations coincided with the manufacturer's: <0.005 mU/L for TSH, <0.30 pmol/L for FT4, and <0.30 nmol/L for T3. Functional sensitivity for TSH was 0.044 mU/L. Over the analytical range tested, within-assay imprecision was below 3.2% for TSH, 2.2% for FT4 and 9.6% for T3, and interassay CVs were below 4.0% for TSH, 5.9% for FT4 and 12.9% for T3. Measurement of diluted sera showed the TSH assay to overestimate recoveries by 18.6%. We have compared sera results of the Elecsys ECLIA assays with those obtained from the IRMA (Spectria-Orion Diagnostica) for TSH: TSH (ECLIA) = 0.074+0.953 TSH (IRMA), (r = 0.974; Sy/x = 2.638), and RIA (Coat a Count-DPC) for FT4:FT4 (ECLIA) = 5.043+0.682 FT4 (RIA), (r = 0.770; Sy/x = 4.774) and RIA (Spectria-Orion Diagnostica) for T3: T3(ECLIA) = -0.461+1.084 T3 (RIA), (r = 0.970; Sy/x = 0.412). When sera from 102 patients were processed by both methods, minimal disagreement in the area of diagnostic classification was observed in 8/102 (7.8%) of the cases. CONCLUSION: The Elecsys 2010 is specially attractive as a routine assay because it is fully automated, obtaining results in only 18 minutes. The analytical assay performance for TSH, FT4 and T3 was shown to be acceptable. Using two different sets of diagnostic tests minimal discrepancies were found in the laboratory assessment for the classification of patients with clinical suspicion of thyroid disease.

Electrochemistry↗

Frequency of oropharyngeal candidiasis in HIV-infected patients on protease inhibitor therapy.

OBJECTIVE: The purpose of this study was to investigate the effect of HIV-1 protease inhibitors on the frequency of oropharyngeal candidiasis in HIV-infected patients. STUDY DESIGN: A clinical and analytic follow-up was carried out to determine the number of episodes of oropharyngeal candidiasis during HIV-1 protease inhibitor therapy and the relation of this incidence to the CD4 lymphocyte count and circulating neutrophils level. Seventy-five HIV-positive patients were selected, and HIV-1 protease inhibitor therapy was administered to each patient over a minimum of 6 months. These patients did not receive long-term preventive antifungal therapy for oral candidiasis, even as secondary prophylaxis against cryptococcosis. Results were compared with those obtained during the previous 6 months, during which patients had been treated only with reverse transcriptase inhibitors. RESULTS: At least one episode of oropharyngeal candidiasis was seen in 56% (42/75) of patients during reverse transcriptase inhibitor therapy and in only 9.3% (7/75) of patients after the initiation of protease inhibitor therapy. The number of relapses decreased significantly when the 2 follow-up periods were compared (P<.0001). The CD4 and CD8 lymphocyte counts increased significantly with protease inhibitor therapy (P<.001 and P<.05, respectively). During reverse transcriptase inhibitor treatment, the probability of the presentation of oropharyngeal candidiasis correlated with falling CD4 counts (P<.0001). The HIV-1 protease inhibitor therapy was associated with a significant increase in the neutrophil count (P<.01). The probability of the occurrence of some episode of candidiasis correlated inversely with the circulating neutrophil level (P<.05). CONCLUSIONS: Protease inhibitor therapy decreases the frequency of HIV-related oropharyngeal candidiasis. The mechanism involved is unknown, but it can be speculated that a reduction of the viral load increases the number of intact T helper cells, which in turn enhances the number of circulating polymorphonuclear neutrophils and regulates their function by means of colony-stimulating factors.

Adult↗

Neurosyphilis in HIV-infected patients.

To determine the prevalence and the clinical and serological findings of neurosyphilis in HIV-infected patients, Treponema pallidum hemagglutination (TPHA) tests, CD4+ lymphocyte counts and determination of rapid plasma reagin (RPR) titers were performed in 972 HIV-infected patients over a period of 3.5 years. Patients were scored according to the Centers for Disease Control's classification for HIV infection. Reactive serum syphilis tests and positive cerebrospinal fluid (CSF)-Venereal Disease Research Laboratory (VDRL) tests, with or without clinical symptoms, were used as the criteria for diagnosis of neurosyphilis. The TPHA test was positive in 31 patients, representing 3.1% of all HIV-infected patients included in the study. Of these, 13 were intravenous drug addicts, 14 were homosexuals and 4 were heterosexuals. Diagnosis of syphilis was concurrent with HIV infection in 19 patients, prior to HIV infection in 6 patients and after HIV infection in 6 patients. CSF examinations were performed in 28 of the 31 (90.3%) patients with serologically evident syphilis. Four patients had positive CSF-VDRL tests with pleocytosis (23.5% of untreated syphilis patients in whom CSF was examined), three of whom reported mild headache, which was considered a doubtful manifestation of neurosyphilis. Patients with syphilis diagnosed and treated prior to diagnosis of HIV infection did not have evidence of neurosyphilis. Seven patients had pleocytosis with a negative CSF-VDRL test, without any clinical manifestations of neurosyphilis. There was no significant difference in the mean CD4+ lymphocyte count between patients with and without neurosyphilis (p = 0.5). RPR titers in neurosyphilis patients were greater than those in patients previously treated for syphilis and in those with pleocytosis only (p = 0.046 and 0.036, respectively). All neurosyphilis patients had an RPR titer > 1:8. After therapy, neurosyphilis patients had negative CSF-VDRL tests with a lower level of pleocytosis. The prevalence of neurosyphilis was 0.4% in HIV-infected patients and 23.5% in HIV-infected patients with untreated syphilis. This high prevalence of neurosyphilis warrants CSF examination in HIV-infected patients with syphilis, regardless of the stage of syphilis.

Adult↗

Changes in the temporomandibular joint caused by the vertical facial pattern. Study on an experimental model.

An experimental model reproducing open bite or verticalized facial pattern was used to study its effect on the temporomandibular joints. 140 Wistar rats were used, divided into 3 groups: bilateral resection of the masseteric muscle, simulated muscular resection and control group. A series of radiological, morphological and histological tests were analyzed. The posterior rotation of the jaw caused by muscular resection although not producing a degenerative effect, did produce specific articular changes in the temporomandibular joint components.

Animals↗

Changes in temporomandibular joint after mandibular subcondylar osteotomy: an experimental study in rats.

The objective of this study was to analyze the changes in the different components of the temporomandibular joint and their relation with age after subcondylar osteotomy. For this purpose 149 Wistar rats were divided into three groups: osteotomy, sham operation, and control. Two experimental ages (30 and 70 days) and radiologic, morphometric, macroscopic, and histologic methods of analysis were used. Different changes were observed in young and adult animals. Subcondylar mandibular osteotomy in growing rats caused anteroinferior displacement of the temporal and mandibular component of the joint. At the end of the experimental period only condylar displacement was maintained. Other changes were flattening of the mandibular condyle, which was transient, and lateral thickening of the articular disc. Subcondylar mandibular osteotomy in adult rats caused slight anterior displacement of the joint components at the end of the experimental period. At this age, although changes similar to those in growing animals were observed in some cases, in other cases the presence of pathologic findings, such as deformation of the condylar cartilage, thickening of the disc, intra-articular adherences, and osteolysis of the temporal fossa, were observed. Changes in the joint components were more marked in growing rats than in adults. In growing rats they affected the form of the condylar process to a greater extent, but the majority of these changes were transient. In adult rats, alterations in the joints were less pronounced, but they affected a greater number of joint components and were permanent.

Adaptation, Physiological↗

Usefulness of urinary growth hormone (GH) measurement for evaluating endogenous GH secretion in acromegaly.

We investigated the relationship between urinary growth hormone (u-GH) and spontaneous 24-hour plasma GH secretion in 15 acromegalic patients. To measure u-GH, we have developed a method based on concentrating the sample by centrifugal ultrafiltration and then performing an immunoradiometric assay using commercially available reagents. u-GH correlated well with the integrated concentration of plasma GH (r = 0.66, p < 0.02). Additionally, u-GH excretion in acromegalic patients was significantly higher than in the control group (190 +/- 100 vs. 3.89 +/- 0.56 pg/min, mean +/- SEM, p < 0.001). Immunoreactive u-GH showed the same elution pattern in Sephadex G-75 as standard or labeled hGH, proving that the substance measured in urine is authentic GH. In conclusion, u-GH appears to be a simple, noninvasive and inexpensive test for evaluating GH secretion in active acromegaly.

Acromegaly↗

[Oro-esophageal candidiasis resistant to fluconazole in patients with AIDS].

BACKGROUND: Fluconazole-resistant Candida albicans has been seldom reported in the literature. in this study, two clinical cases of oro-esophageal candidosis resistant to this antifungal drug are described. METHOD: Infection was confirmed by isolation of yeasts (culture of mouth swabs, oesophagoscopy and histology of biopsy specimens). In vitro antifungal susceptibility tests were performed with standard methods (NCCLS). RESULTS: Clinical and microbiological fluconazole-resistant Candida albicans was isolated. CONCLUSIONS: Causes of development of resistances are commented, and the role of immunodeficiency and the possibility of a Candida strain selection is discussed.

AIDS-Related Opportunistic Infections↗

Osteochondroma induced by reflection of the perichondrial ring in young rat radii.

In order to investigate the effects of reflection of the perichondrial ring in osteochondroma formation, the perichondrial rings of the epiphyseal growth plates in 42 young rat radii were turned to the metaphyseal periosteum by means of blunt dissection. Seven days after surgery a small nest of chondrocytes appeared on the metaphyseal-diaphyseal bone surface at the level of the tip of the reflected perichondrial ring. From the 9th to the 15th days the histological pattern of the osteochondroma was established. The osteochondroma was not connected with the hypertrophic cartilage of the growth plate. During the third and fourth weeks the osteochondroma began to regress with the disappearance of the cartilage nest. During the development of the lesion the bone grew normally and the growth plate migrated distally while the lesion remained at its initial site. The growth plate zone devoid of perichondrial ring was covered by fibrous connective tissue and no removal of the perichondrial ring occurred. These results suggest that the origin of this osteochondroma is the perichondrial ring cells whose polarity has been surgically changed.

Animals↗