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Biomedical subjects

M Hermida

Publications and source records attributed to M Hermida.

At least 19 recordsLinked to original sources

Detection of HCV-RNA in saliva of HIV-HCV coinfected patients.

The presence of HCV-RNA in saliva of patients with chronic hepatitis C provides a biological basis for the potential transmission of this virus. HCV viremia is particularly high in HCV-HIV-coinfected patients, which could favor the presence of HCV in their saliva. This study was designed to evaluate the prevalence of HCV in saliva of HCV-HIV-coinfected patients. Stimulated whole saliva was collected from 75 HCV-HIV-coinfected patients and 75 HCV controls. The presence of HCV-RNA in saliva was tested by a highly sensitive noncommercialized nested PCR, and analyzed in relation to demographic, clinical, and analytical variables. HCVRNA was detected in the saliva of 49 (65%) HCV-HIV-coinfected patients and 39 (52%) HCV controls. The presence of HCV in saliva was not related to any of the analyzed variables in HCV-HIV-coinfected patients. In the HCV control group a statistically significant relationship was demonstrated only between the detection of HCV-RNA in saliva and the viral load in peripheral blood (p < 0.001). Our results indicate that there is a trend toward a higher HCV-RNA prevalence in the saliva of HCV-HIV-coinfected patients.

Adult↗

HCV clearance patterns in saliva and serum of patients with chronic HCV infection under interferon plus ribavirin therapy.

STATEMENTS OF THE PROBLEM: Hepatitis C virus (HCV)-RNA is often present in saliva of HCV-infected patients, with plasma viral load being the only known predictable factor. Interferon plus ribavirin therapy yields a sustained reduction in HCV viremia. This study aimed to assess the presence of HCV in saliva and serum specimens from patients undergoing this combination therapy (CT). METHOD OF STUDY: Paired serum and saliva specimens were collected from 44 chronic HCV-infected patients at basal time, 4 and 12 weeks after CT onset, at the end of treatment and 6 months latter. Serum HCV-RNA levels were determined by the polymerase chain reaction (PCR) Amplicor system. Presence of HCV-RNA in saliva was tested by a highly sensitive non-commercialized nested-PCR. RESULTS: The HCV-RNA was detected in 26 saliva specimens at basal time (59.1%). In 34.1% of cases, a concordance viral clearance pattern in serum and saliva was observed in both responders (pattern 1a) and non-responders (pattern 1b). In pattern 2 (13.6% of cases), HCV was detected longer during CT in serum than in saliva (pattern 2a) or in saliva than in serum (pattern 2b). In 11.3% of patients, viral clearance was corroborated either in their serum (pattern 3a) or in their saliva (pattern 3b), but not in both fluids. Of the eight primary responders with 1a clearance pattern, seven were sustained responders. None of the patients with 2a clearance pattern was a sustained responder. Of the two primary responders showing the 3b salivary pattern, one had already relapsed in the first 6 months of follow up. CONCLUSIONS: The present results suggest that the monitoring of salivary levels of HCV would be a helpful means of determining sustained antiviral effects of interferon and ribavirin in the treatment of HCV disease.

Adolescent↗

[GB virus C: lack of association with transaminases levels, CD4 and HIV viral load in aids patients].

OBJECTIVE: To study the prevalence of GBV-C-RNA in sera of HIV-infected patients and determine whether differences in immunological condition and hepatic disease exist between GBV-C positive and negative patients. METHODS: The presence of GBV-C-RNA was determined in sera of 222 HIV-positive patients by semi-automated RT-PCR. A comparison of GBV-C-RNA positive and negative patients was made by studying a series of clinical and analytical parameters. This same comparison was made in particular between those coinfected with HCV and GBV-C and those who only presented GBV-C. RESULTS: Prevalence of GBV-C-RNA was 28.8%. The most frequent hepatotropic virus was HCV, appearing in 71.6% of cases. Coinfection with HCV and HGV was present in 17% and 8.6% only had GBV-C. Patients positive for GBV-C-RNA showed clinical and analytical characteristics similar to those found in GBV-C-RNA negative patients. Among the HCV-GBV-C coinfected and those presenting HGV as the only virus it was observed that the coinfected group presented alterations in transaminases and predominance of parenteral transmission as a risk factor for HIV, whereas the GBV-C group presented normal transaminases and predominance of sexual transmission. No differences were perceived in mean CD4 and HIV-RNA values in both groups. CONCLUSIONS: Being positive for GBV-C in HIV-positive patients does not influence the presence of hepatic disease that in these patients is frequently accompanied by coinfection with other hepatotropic viruses. Moreover, it does not seem to influence the viremia of the HIV nor the CD4 cell counts.

Adolescent↗

Determination of polarimetric parameters of honey by near-infrared transflectance spectroscopy.

NIR transflectance spectroscopy was used to determine polarimetric parameters (direct polarization, polarization after inversion, specific rotation in dry matter, and polarization due to nonmonosaccharides) and sucrose in honey. In total, 156 honey samples were collected during 1992 (45 samples), 1995 (56 samples), and 1996 (55 samples). Samples were analyzed by NIR spectroscopy and polarimetric methods. Calibration (118 samples) and validation (38 samples) sets were made up; honeys from the three years were included in both sets. Calibrations were performed by modified partial least-squares regression and scatter correction by standard normal variation and detrend methods. For direct polarization, polarization after inversion, specific rotation in dry matter, and polarization due to nonmonosaccharides, good statistics (bias, SEV, and R(2)) were obtained for the validation set, and no statistically (p = 0.05) significant differences were found between instrumental and polarimetric methods for these parameters. Statistical data for sucrose were not as good as those of the other parameters. Therefore, NIR spectroscopy is not an effective method for quantitative analysis of sucrose in these honey samples. However, NIR spectroscopy may be an acceptable method for semiquantitative evaluation of sucrose for honeys, such as those in our study, containing up to 3% of sucrose. Further work is necessary to validate the uncertainty at higher levels.

Calibration↗

Detection of HCV RNA in saliva of patients with hepatitis C virus infection by using a highly sensitive test.

Hepatitis C virus (HCV) is transmitted primarily by direct percutaneous exposures to blood. Since HCV RNA has been found in saliva, it has been suggested that saliva might also be a source of infection. HCV RNA in saliva from plasma HCV RNA positive patients was tested by a highly sensitive PCR method. HCV RNA was detected in 32 out of 61 saliva specimens (52.4%). No correlation was found between the presence of HCV in saliva and age, sex, identified risk factors for HCV infection, time lapsed since the diagnosis, transaminases and alkaline phosphatase values and stimulated salivary flow. A statistically significant relation between plasma HCV RNA viral load and saliva HCV RNA detection was observed (P<0.001). In conclusion, HCV RNA is often present in saliva of HCV infected patients, with plasma viral load being the only known predictable factor. Further studies on salivary HCV RNA are needed.

Adult↗

Synaptonemal complex analysis in oocytes and spermatocytes of threespine stickleback Gasterosteus aculeatus (Teleostei, Gasterosteidae).

A surface-spreading synaptonemal complex (SC) technique was employed to analyze spermatocytes and oocytes of stickleback, Gasterosteus aculeatus, in order to visualize the process of chromosome synapsis. The mean SC length was 150 +/- 18 microm in three males and 143 +/- 12 microm in one female analyzed. A representative SC karyotype with 21 bivalents was also presented. Each SC had lateral elements of equal length. No bivalent displaying the atypical synaptic behaviour which is often associated with heteromorphic sex chromosomes was observed neither in males nor in the female analyzed.

Animals↗

Removal of bowel aerobic gram-negative bacteria is more effective than immunosuppression with cyclophosphamide and steroids to decrease natural alpha-galactosyl IgG antibodies.

Natural alpha-Galactosyl (Gal) antibodies play an important role in the rejection of pig xenografts by humans and Old World monkeys. In this study we investigate the efficacy of two different strategies to reduce the serum level of natural anti-Gal antibodies. On the one hand, removal of aerobic gram-negative bacteria from the intestinal flora, because anti-Gal antibodies appear to be produced as a result of the continuous sensitization by these microorganisms. On the other hand, we studied the effect on these antibodies of an immunosuppressive regimen of cyclophosphamide and steroids. Ten baboons were treated for three months with norfloxacin (Nor Group; n=6) or cyclophosphamide and steroids (CyP Group; n=4). A further four baboons did not receive any treatment (Control Group). Aerobic gram-negative bacteria became negative in stools of the Nor Group after two weeks of treatment, and remained undetectable until week 7. Thereafter, a gradual increase on the fecal concentration of aerobic gram-negative bacteria was observed despite the norfloxacin treatment. The mean anti-Gal IgG in the Nor Group gradually declined from week 4 to 9 to a mean of 62.7 +/- 18% of the baseline level, and during this period were significantly lower than in the CyP (P<0.02) and the Control (P<0.05) groups. No differences were observed between the three groups during the 16 weeks of follow-up in serum levels of anti-Gal IgM, hemolytic anti-pig antibodies, total IgG, IgM and IgA. In conclusion, removal of normal aerobic gram-negative bacteria from the intestinal flora is more effective than immunosuppression with CyP and steroids in reducing the level of natural anti-Gal antibodies, although there is no discernible effect on IgM antibodies.

Animals↗

Major components of honey analysis by near-infrared transflectance spectroscopy.

NIR transflectance spectroscopy was used to analyze fructose, glucose, and moisture in honey. A total of 161 honey samples were collected during 1992 (46), 1995 (58), and 1996 (57). Samples were analyzed by instrumental, enzymatic (fructose and glucose), and refractometric (moisture) methods. Initially, different calibrations were performed for each of the 3 years of sampling. Good predictions were obtained for all three components with equations of the particular year. But good predictions were not always obtained when the equations calculated one year were applied to samples from another year. To perform a lasting calibration, unique calibration (121 samples) and validation (40 samples) sets were built; honeys of the 3 years were included in both sets. Good statistics (bias, standard error of validation (SEV), and R(2)) were obtained for all three components of the validation set. No statistically significant differences (p = 0.05) were found between instrumental and reference methods.

Calibration↗

Effects of the addition of Enterococcus faecalis in Cebreiro cheese manufacture.

Ten batches of Cebreiro, a fresh or short-ripened acid-curd cheese, produced in the Galician mountains (NW Spain) were prepared from pasteurized milk inoculated with microorganisms isolated from raw-milk cheese. Two control batches were made with a Lactococcus lactis subsp. lactis starter; 8 batches were made with the lactococcal starter plus one of eight Enterococcus faecalis cultures: 4 E. faecalis var. liquefaciens (EFLB) and 4 E. faecalis var. faecalis (EFFB). Whey dry matter in the EFLB was notably higher than in the control batches and this was related to lower cheese yields. After over 15 days storage the highest counts of both aerobic bacteria and lactic acid bacteria were observed for the EFLB. The lower content in protein on dry matter was found in the EFLB. The beta-casein broke down to a greater extent in the EFLB than in the EFFB, the lowest values being obtained for the control batches. The higher level of hydrolysis of alphax1-casein and maximum peptide alpha(s1) - I/alpha(s1)-casein ratio were obtained for EFFB at day 15 of storage. In all the batches made with enterococci soluble nitrogen was higher than in the control batches, with the highest values in the EFLB. In all the batches made with enterococci, volatile free fatty acid, long-chain free fatty acids and diacetyl and acetoin contents at days 10 and 15 of storage were higher than in the control batches, the highest values being obtained for EFLB. Acetic acid in all batches accounted for the main proportion of the volatile free fatty acids. Butyric and caproic acids were not detected in the volatile free fatty acids fractions of the control batches, but both acids were detected in most of the batches made with enterococci. The more intense acid taste was found in the EFFB and control batches, the most bitter taste being found in the EFLB. Buttery, rancid and spicy flavors were more evident in the EFLB. The rancid and spicy flavors were positively correlated with the contents of volatile free fatty acids and long-chain free fatty acids. The cheeses of EFLB proved to be more crumbly than the EFFB, whereas the stickiness and deformability were higher in the EFFB. The batches with similar organoleptic characteristics to those of traditional cheese were the batch IV made with the less proteolytic strain of E. faecalis var. liquefaciens, and the batch VI made with a moderate lipolytic activity strain of E. faecalis var. faecalis.

Animals↗

Long-term re-treatment with interferon and ribavirin combination therapy in patients with chronic hepatitis C who are non-responders to interferon alone: a preliminary study.

We evaluated the efficacy and tolerance of ribavirin and IFN-alpha combination therapy over 12 months in 28 patients who were non-responders to IFN-alpha alone. Of 24 patients who have finished the therapy, 6 (25%) obtained a complete response (normal ALT and negative HCV RNA) at the end of treatment and maintained a sustained response 27% (5/18).

Adult↗

[The efficacy of interferon alfa treatment in chronic hepatitis C in relation to the serum concentration of RNA-HCV and the viral genotype].

BACKGROUND: To determinate the viral variables (viremia and genotype) and patients variables (route and years of infection, histology, etc) related with response to the treatment with alpha intefferon in patients with chronic hepatitis C. SUBJECTS AND METHODS: We have studied 50 patients with chronic hepatitis C that received an intefferon treatment during a year. In all them it was accomplished a hepatic biopsy before of the beginning of treatment and it was studied the viral load by quantitative PCR (Monitor, Roche) at the beginning of treatment, at four weeks and at the end of treatment. It was determined the HCV genotype by method INNO-LIPA (Boehringer Inghelheim). RESULTS: 8 patients (16%) achieved a complete sustained response during all the follow-up period (13.8 + 1.6 months); 8 patients responded but relapsed when the intefferon was interrupted and 34 patients not responded. The responders had lower viremia (p < 0.032) and HCV RNA negative at the fourth treatment week (p < 0.0082). The genotype 1b was the most frequent, the one which was presenting a smaller percentage of response and it was associated with a higher viremia. Cirrhotic patients were the worse responders. Ex-lDAs were better responders than the others groups, however this difference did not show statistics meaning. CONCLUSIONS: A low pretreatment HCV-RNA level seems to be indicative of a sustained response to IFN, Whereas a high level seems to be indicative of a non sustained response to IFN. Negativization of HCV-RNA at week four is a good indicator of response to intefferon. Genotype 1b is the most frequent and the worse responder. To have 1b genotype, high pretreatment HCV-RNA level and to be viremic at four treatment seems to be indicative of non response.

Adult↗

Characterization of penicillinase-producing strains of Neisseria gonorrhoeae.

Twenty-three penicillinase-producing strains of Neisseria gonorrhoeae were characterized. All strains showed hybridization with a TEM-1 probe and production of a TEM-1 type beta-lactamase (pI 5.4) on isoelectric focusing. Eight strains also showed a beta-lactamase band of pI 5.5. The penicillin MICs for the strains producing only TEM-1 were in the range 4-16 micrograms/ml; MICs of the strains with the additional pI 5.5 band were > or = 128 micrograms/ml for seven strains and 4 micrograms/ml for an arginine-hypoxanthine dependent strain. The association of the high MICs with the presence of the pI 5.5 band was statistically significant. The pI 5.5 band could represent a new beta-lactamase type or a TEM-1 mutant.

Isoelectric Point↗

A survey of beta-lactamases from 618 isolates of Klebsiella spp.

beta-Lactamases from 618 isolates of Klebsiella spp. (487 Klebsiella pneumoniae and 131 Klebsiella oxytoca) were identified by analytical isoelectric focusing. The strains were consecutive isolates from 618 different patients in five hospital centres between 1980 and 1984. Most of the chromosomal beta-lactamases from the K. pneumoniae isolates had a pI of 7.1, and most of the K. oxytoca isolates had chromosomal beta-lactamases of pIs 7.8, 5.3 and 5.5. Plasmid-mediated beta-lactamases were produced by 91.8% of K. pneumoniae isolates and 9.9% of K. oxytoca isolates. The beta-lactamase types identified most frequently were SHV-1 in the former species, and TEM-1 in the latter species. Of the K. pneumoniae isolates, 101 produced both of these plasmid-mediated beta-lactamases, while one strain produced an extended-spectrum SHV-like beta-lactamase. Of the 131 K. oxytoca isolates, 20 were hyper-producers of chromosomal beta-lactamase. The susceptibility of this genus to beta-lactam antibiotics is controlled almost exclusively by beta-lactamases, the production of which may differ from one strain to another; thus the empirical therapeutic use of beta-lactam antibiotics is not recommended for this genus.

Chromosomes, Bacterial↗

[Characterization of Haemophilus influenzae's resistance to ampicillin].

BACKGROUND: The resistance of Haemophilus influenzae to ampicillin is nearly always dependent on plasmid-mediated betalactamase production. The betalactamase type most frequently identified was TEM-1; type ROB has occasionally been identified. The presence of plasmid-mediated betalactamase has been studied in H. influenzae strains moderately susceptible or resistant to ampicillin. METHODS: The test used in the detection of the presence of betalactamase was nitrocefin hydrolysis; the enzyme type was identified by analytic isoelectric focusing; the codifying gene of this betalactamase was identified by hybridisation with a TEM-1 probe labeled with digoxigenine. Strains studied were 110, obtained during the period 1987 to 1989 and isolated from four Hospitals of the Vallés area of Barcelona. RESULTS: Enzyme production was detected in 105 of 110 strains studied; TEM-1 type betalactamase was identified in 108 and positive hybridisation was obtained in these strains with the TEM-1 probe. CONCLUSIONS: Nitrocefin hydrolysis is a reliable test for the detection of betalactamase production, although it may be unable to detect it (falses negatives) when the production is low. The only betalactamase type identified was TEM-1. The frequency of the strains with non-enzymatic resistance was 1.8%.

Adult↗

[Comparative study with digoxigenin and biotin markers for the DNA probe identification of genes coding for type TEM beta-lactamases].

We have tested two non-isotopic labels: digoxigenin--11-dUTP and biotin-7-dATP for the detection of TEM-1 beta-lactamase gene with a TEM-1 probe by DNA:DNA hybridisation (using spot technique). The use of radioactive labels is inconvenient and not available to all the clinical bacteriological laboratories. The strains tested were: 16 on file (14 carriers of different types of beta-lactamase) and 19 clinical isolated strains. Previously, the plasmid beta-lactamase of these strains had been identified by analytical isoelectric focusing. Using the probe labeled with digoxigenin-11-dUTP, the correlation between results obtained by DNA:DNA hybridisation and those by analytical isoelectric focusing was excellent, with no false positives; with the biotin--7-dATP labeled probe, reading was difficult and some false positives were observed.

Biotin↗

Activity of amoxycillin/clavulanate against beta-lactamase-producing Escherichia coli and Klebsiella spp.

We studied the susceptibility to the amoxycillin/clavulanate combination (2:1), and the characteristics of the beta-lactamases, of 1410 amoxycillin-resistant strains of Enterobacteriaceae (785 of Escherichia coli and 625 of Klebsiella spp.). Of the E. coli strains, 97.07% were susceptible to the combination and 98.85% produced plasmid-mediated beta-lactamases. Of the Klebsiella spp. strains, 87.84% were susceptible to the combination and 75.36% produced plasmid-mediated beta-lactamases. In both cases, the differences in MIC values, according to the characteristics of the beta-lactamases of the strains, were statistically significant. Of the 99 strains of E. coli and Klebsiella spp. that were not inhibited by amoxycillin 16 mg/l with clavulanate 8 mg/l, 19 hyperproduced chromosomal beta-lactamase; five produced a plasmid-type beta-lactamase as well. Fifty-seven strains produced two plasmid-type beta-lactamases.

Amoxicillin↗