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Biomedical subjects

M Bosch

Publications and source records attributed to M Bosch.

At least 19 recordsLinked to original sources
PubMed2006

Single-nucleotide polymorphisms in the epsilon-globin gene for differentiating primate infraorders.

Molecular methods are being used to enforce wildlife conservation laws by identifying the species or the geographic origin of an unknown sample. However, the promising use of single-nucleotide polymorphisms (SNPs) in this field is still widely unexplored. In the present work, we have developed a reliable and easy method based on single-base extension technology for the scoring of 3 SNPs in the epsilon-globin gene that successfully identifies the primate infraorder a sample belongs to. Since primates are of high conservation concern and different infraorders are distributed in specific parts of the world, this method will serve for an initial potentially automated screening of the taxonomy and geographic origin of an unknown primate sample arriving at customs.

Animals↗
PubMed2006

[In vitro activity of voriconazole against yeast and algae isolates according to new resistance pattern cut-off points].

Voriconazole is a second-generation triazole derived from fluconazole but with greater potency and spectrum of activity, showing good in vitro activity against Candida, Cryptococcus and Aspergillus species, and other filamentous and dimorphic fungi. It can be administered orally or intravenously. It was initially approved in 2002 by the U.S. Food and Drug Administration as a treatment option for invasive aspergillosis and Fusarium and S. apiospermum infections showing resistance or intolerance to other antifungals; later on, it also received approval in the United States and Europe as a treatment option for esophageal candidiasis; candida infection in non-neutropenic patients; disseminated candidiasis of skin, abdomen, kidney and bladder; and injuries. Recently, the Clinical Laboratory Standard Institute established some provisional break points for voriconazole, classifying isolates with an MIC or=4 mg/l as resistant. In line with these new data, we performed a systematic review of literature on in vitro activity of voriconazole against yeast and algae isolates, and compared it to that of fluconazole and itraconazole. The review included a total of 27,340 yeast isolates, 24,177 of Candida species, 2,726 of Cryptococcus species, 453 of other species, and 104 Prototheca. The yeast isolates resistant to voriconazole is approximately 1%, and 71% of fluconazole-resistant isolates are susceptible to voriconazole.

Antifungal Agents↗
PubMed2006

[Evaluation and utility of the E-test and Neo-Sensitabs methods in studying fluconazole yeast susceptibility].

Standardized broth dilution methods are cumbersome for routine use in a clinical laboratory to study antifungal yeast susceptibility. Recently, the CLSI has standardized a disk diffusion method faster and more suitable to study fluconazole and voriconazole susceptibility. The objectives of the present study were to determine: a) the suitability of the Neo-Sensitabs tablets to study fluconazole susceptibility; b) whether Mueller-Hinton agar with methylene blue (MHAG-AM) could be used in the E-test method; and c) the interaction of the methylene blue with RPMI medium. A total of 84 blood stream yeast isolates were used (25 C. albicans, 7 C. parapsilosis, 10 C. tropicalis, 12 C. glabrata, 7 C. krusei, 4 C. lusitaniae and 19 C. neoformans). The methylene blue makes sharper inhibition zones both in MHAG-AM and RPMI media. With fluconazole Neo-Sensitabs tablets, the lowest percentage of very major errors was found in MHAG-AM and the greatest in RPMIG. In both diffusion methods and culture media, the very major errors were found in C. albicans, C. tropicalis (only with Neo-Sensitabs) and C. glabrata. The percentage of fluconazole-resistant strains was lower in the media that contained glucose (2%). Neo-Sensitabs tablets are a reliable alternative to the dilution methods to detect fluconazole susceptibility. In the case of resistance, more studies are required; nevertheless, inhibition zone > or =17 mm should be applied to define fluconazole resistance.

Antifungal Agents↗
PubMed2005

Circulating IL-18 concentration is associated with insulin sensitivity and glucose tolerance through increased fat-free mass.

AIMS/HYPOTHESIS: Knowledge of the factors which simultaneously contribute to insulin-resistance-related inflammation may contribute to early therapeutic targeting. IL-18 has recently been described as one of the factors which, in addition to insulin resistance, may also contribute to atherosclerosis. However, the source of IL-18 is not well characterised. MATERIALS AND METHODS: We aimed to study body composition (bioelectric impedance), glucose tolerance (OGTT) and insulin sensitivity (minimal model method) in relation to serum IL-18 (ELISA) concentration in 144 otherwise healthy men aged 51.9+/-12.5 years. RESULTS: In contrast to previous observations in women, circulating IL-18 was not significantly associated with BMI (r=0.12, p=0.1) or WHR (r=0.08, p=0.3). IL-18 was also not associated with absolute or percent fat mass (bioelectric impedance, p>0.20) but, interestingly, it was significantly linked to fat-free mass (p=0.03). Serum IL-18 increased with each quartile of fat-free mass, corresponding to values of < or = 64.2; >64.2 to < or = 71.6; >71.6 to < or = 80.9; and > or = 80.9 kg (ANOVA, p<0.0001). IL-18 was more closely associated with postload glucose during an OGTT (p=0.04) rather than with fasting glucose (p=0.1). HbA1c (p=0.03), HDL-cholesterol (p=0.04) and serum triglycerides (p=0.03) and parameters of systemic inflammation (C-reactive protein, p=0.02) were also significantly associated with circulating IL-18. Insulin sensitivity (minimal model analysis) was linked to circulating IL-18 (p=0.01). In a multiple linear regression analysis this relationship remained significant after controlling for BMI, age and glucose tolerance status. In another model, both fat-free mass and insulin sensitivity contributed to 10% of IL-18 variance. CONCLUSIONS/INTERPRETATION: Fat mass does not seem to influence circulating IL-18, as initially proposed. In contrast, the fat-free mass compartment (a well-known confounder in the evaluation of insulin sensitivity) may significantly contribute to the relationship between IL-18 and insulin action.

Adipose Tissue↗
PubMed2005

Long-term expression of erythropoietin from myoblasts immobilized in biocompatible and neovascularized microcapsules.

The present paper investigates the long-term functionality of an ex vivo gene therapy approach based on cell microencapsulation for the continuous delivery of erythropoietin (EPO) without implementation of immunosuppressive protocols. Polymer microcapsules (0.5 ml) loaded with EPO-secreting C(2)C(12) myoblasts and releasing 15,490 +/- 600 IU EPO/24 h were implanted in the peritoneum and subcutaneous tissue of syngeneic and allogeneic mice. High and constant hematocrit levels were maintained for more than 100 days in all implanted mice. Capsules retrieved from the peritoneum were free-floating or forming small capsule clusters, and we detected only a weak fibroblast outgrowth in capsules adhered to organs, whereas capsules explanted from the subcutaneous region appeared altogether as a richly vascularized structure with no signs of major host reaction. Interestingly, the functionality of capsules implanted in the allogeneic mice persisted until day 210 after implantation. These results highlight the feasibility of cell encapsulation technology for the long-term delivery of EPO independent of the method of administration and the mouse strain.

Animals↗
PubMed2005

Frequency and distribution of chromosome abnormalities in human spermatozoa.

This study reviews the frequency and distribution of numerical and structural chromosomal abnormalities in spermatozoa from normal men obtained by the human-hamster system and by multicolor-FISH analysis on decondensed sperm nuclei. Results from large sperm karyotyping series analyzed by chromosome banding techniques and results from multicolor FISH in sperm nuclei (of at least 10(4) spermatozoa per donor and per probe) were reviewed in order to establish baseline values of the sperm chromosome abnormalities in normal men. In karyotyping studies, the mean disomy frequency in human sperm is 0.03% for each of the autosomes, and 0.11% for the sex chromosomes, lower than those reported in sperm nuclei by FISH studies using a similar methodology (0.09% and 0.26%, respectively). Both types of studies coincide in that chromosome 21 and sex chromosomes have a greater tendency to suffer segregation errors than the rest of the autosomes. The mean incidence of diploidy, only available from multicolor FISH in sperm nuclei, is 0.19%. Inter-donor differences observed for disomy and diploidy frequencies among FISH studies of decondensed sperm nuclei using a similar methodology could reflect real differences among normal men, but they could also reflect the subjective application of the scoring criteria among laboratories. The mean frequency of structural aberrations in sperm karyotypes is 6.6%, including all chromosome types of abnormalities. Chromosome 9 shows a high susceptibility to be broken and 50% of the breakpoints are located in 9q, between the centromere and the 9qh+ region. Structural chromosome aberrations for chromosomes 1 and 9 have also been analyzed in human sperm nuclei by multicolor FISH. Unfortunately, this assay does not allow to determine the specific type of structural aberrations observed in sperm nuclei. An association between advancing donor age and increased frequency of numerical and structural chromosome abnormalities has been reported in spermatozoa of normal men.

Adult↗
PubMed2005

[Activity of voriconazole against yeasts isolated from blood culture determined by two methods].

The in vitro activity of voriconazole has been determined by two methods: the reference M27-A2 and the marketed Sensititre YeastOne microdilution colorimetric method. The agreement (+/-2 dilutions) and correlation between methods as well as the percentage of errors has been determined. A total of 144 yeasts (47 Candida albicans, 52 C. parapsilosis, 13 C. tropicalis, 10 C. krusei, 9 C. glabrata, 2 C. guilliermondii, 1 C. colliculosa, 1 C. dubliniensis, 2 Trichosporum asahii, 1 T. mucoide, 1 Trichosporum spp., 1 Kloakera apis, 2 Pichia ohmeri, and 2 Rhodotorula glutinis) isolated from blood culture between October 2002 and May 2005 were assayed. Voriconazole has shown good in vitro activity. The rate of voriconazole-susceptible (MIC < or =1 mg/l) strains was 97% and the MIC90 0.25 mg/l by the two methods. The overall percentage of agreement between methods was 86% (range 44.23-100%) and the Pearson's coefficient of correlation was 0.961. Categorical agreement was strain dependent and ranged from 84.6% for emergent yeasts to 100% for the other species tested except for C. glabrata (66.6%). No major or very major errors were found, the percentage of minor errors being 1.38%. Only one C. tropicalis and one C. glabrata strain were resistant (MIC > or =4 mg/l) to voriconazole (1.38%) by the reference method. The colorimetric method identified the voriconazole-resistant C. tropicalis strain, and classified the C. glabrata as susceptible-dose dependent. The colorimetric method is a potential alternative method for testing the susceptibility of yeast in a clinical laboratory and identifies the susceptible strains (100% agreement) very well. Nevertheless, further studies including more voriconazole-resistant strains are required to determine the ability of the method to identify resistance, which is the goal of susceptibility tests.

Antifungal Agents↗
PubMed2004

First metacarpal lengthening following traumatic amputation of the thumb: long-term follow-up.

Thumb amputation is a relatively frequent injury in industrial accidents, producing a significant effect on hand function. The long-term results of 18 first metacarpal elongations in patients who had had a non-replantable thumb amputation at the metacarpophalangeal level are reported. Lengthening was performed using a unilateral external fixator. After a learning curve with a high complication rate and prolonged treatment, good functional results were obtained after some technical modifications, with eight patients returning to their previous occupation, and the rest requiring a change in their job.

Adolescent↗
PubMed2002

[Recovery of lung function after laparoscopic cholecystectomy: the role of postoperative pain].

OBJECTIVES: Lung function has been shown to deteriorate after laparoscopic cholecystectomy (LC). The present study evaluated 1) the rate of recovery after LC, and 2) the pathogenic role of postoperative pain in functional deterioration. DESIGN: Lung function was measured 24 hours before LC, upon hospital discharge (48-72 h after LC), and 10 days later. All patients received metamizol after LC until discharge (2 g every 6 h i.v.). Half the patients (analgesia group) received tramadol (150 mg i.m.) 30 minutes before lung function testing on the day of hospital discharge. The remaining patients constituted the control group. PATIENTS: Twenty healthy subjects (53 4 years old) undergoing LC for gall bladder removal. All signed informed consent forms. Measures and outcomes: Patient characteristics and preoperative lung function results were similar in both groups. LC duration and postoperative course were also similar in both groups. All were discharged without complications within 72 hours after LC. Lung function upon discharge (FVC, FEV1, TLC, PaO2 and AaPO2) had deteriorated in both groups (p<0.001). Deterioration was less marked in the analgesia group (p < 0.05). Ten days later, lung function had normalized for all subjects. CONCLUSIONS: These results indicate that after LC, 1) lung function is still abnormal when the patient is discharged from hospital, 2) lung function has fully recovered within 10 days, and 3) postoperative pain contributes significantly to temporary deterioration in lung function.

Adult↗
PubMed2002

Stabilization of human pancreatic ribonuclease through mutation at its N-terminal edge.

Enzyme stability can be an important parameter in the design of recombinant toxins because unstable proteins are often degraded before they can reach their cellular target. There is great interest in the design of human pancreatic ribonuclease variants that could be cytotoxic against tumoral cells. To this end, some residues in the protein need to be substituted, but this may result in a loss of stability. Previous papers have reported the production of N- and C-terminal human pancreatic ribonuclease variants with increased thermal stability. Here, we investigated the contribution of the different amino acid changes at the N-terminus of the protein to its thermostability increase. We show that this increase correlates with the helical propensity of the first alpha-helix of the protein. On the other hand, deletion of the four last residues of the protein does not affect its thermal stability. These results set the basis for the design of a human pancreatic ribonuclease template on which amino acid substitutions can be made that could render the enzyme cytotoxic, without an important loss in its stability.

Enzyme Stability↗
PubMed2001

Expression of the Pasteurella multocida ompH gene is negatively regulated by the Fur protein.

The fur gene of Pasteurella multocida has been cloned by complementation of an Escherichia coli fur mutant. The P. multocida fur gene, which encodes a predicted protein of 147 amino acids, displaying the highest identity (89%) with the same protein of Haemophilus influenzae, is negatively regulated by its own product. By construction of a P. multocida fur mutant, it has been demonstrated that the ompH gene, encoding a major structural protein of the outer membrane, presenting high antigenicity power, is negatively regulated by iron and glucose. Furthermore, wild-type and fur-defective cells of P. multocida show the same level of virulence.

Animals↗
PubMed2001

Treatment with continuous positive airway pressure is not effective in patients with sleep apnea but no daytime sleepiness. a randomized, controlled trial.

BACKGROUND: The sleep apnea-hypopnea syndrome is defined by a pathologic number of respiratory events during sleep (the apnea-hypopnea index, defined as the number of apnea and hypopnea episodes per hour) and daytime symptoms (mostly, excessive sleepiness). In patients with the sleep apnea syndrome, treatment with continuous positive airway pressure (CPAP) normalizes both the apnea-hypopnea index and diurnal symptoms. However, the effect of CPAP in persons with a pathologic apnea-hypopnea index without daytime sleepiness is unclear. OBJECTIVE: To investigate the short-term effects of CPAP on quality of life, objective sleepiness, cognitive function, and arterial blood pressure in nonsleepy patients with a pathologic apnea-hypopnea index. DESIGN: Multicenter randomized, placebo-controlled, parallel-group study. SETTING: Six teaching hospitals in Spain. PATIENTS: 55 patients with an apnea-hypopnea index of 30 or greater who did not have daytime sleepiness (Epworth Sleepiness Scale score </= 10). INTERVENTION: Patients were randomly assigned to receive optimal (n = 29) or sham (n = 25) CPAP and were observed for 6 weeks. MEASUREMENTS: Quality of life, objective sleepiness (Multiple Sleep Latency Test score), cognitive function, and arterial blood pressure. RESULTS: The intervention and control groups were similar in terms of mean (+/-SE) age (54 +/- 2 vs. 52 +/- 2 years), apnea-hypopnea index (54 +/- 3 vs. 57 +/- 4), Epworth Sleepiness Scale score (7.0 +/- 0.4 vs. 7.0 +/- 0.4) and adherence to CPAP treatment (5.0 +/- 0.4 vs. 4.0 +/- 0.5 hours/d). Other variables, such as quality of life, cognitive function, and arterial blood pressure, were also similar in both groups before treatment. After 6 weeks of CPAP or sham CPAP, none of these variables changed significantly. CONCLUSION: In patients with an apnea-hypopnea index of 30 or greater and no subjective daytime sleepiness, CPAP does not modify quality of life, objective sleepiness, vigilance, attention, memory, information processing, visuomotor coordination, or arterial blood pressure. Treatment with CPAP is therefore not indicated in nonsleepy patients with a pathologic apnea-hypopnea index.

Blood Pressure↗
PubMed2001

Anti-infectious properties of the human milk fat globule membrane.

Secretory immunoglobulin A (sIgA), the predominant antibody fraction of human milk, represents a major protective factor against neonatal infection. Until now, sIgA had been identified only in the humoral fraction of human milk. For bovine milk an association between sIgA and the milk fat globule (MFG) membranes has been demonstrated. The aim of our study was to assess whether sIgA is associated with the MFG membranes in human milk. Using anti-sIgA-agglutinated human MFG and immune fluorescence microscopy, we demonstrated that sIgA is, in fact, associated with human MFG. Subsequently, by electrophoretic separation of human MFG membranes and Western blotting, we demonstrated specific sIgA bands, suggesting that sIgA is truly an integral part of the human MFG membrane. This may be of physiological relevance, as undigested and functional human MFG are found in the stools of the newborn.

Anti-Infective Agents↗
PubMed2001

Acrocentric chromosome disomy is increased in spermatozoa from fathers of Turner syndrome patients.

The aim of the present study was to investigate whether there was an increase of aneuploidy in the sperm from fathers of Turner syndrome patients of paternal origin who, in a previous study, showed an elevated incidence of XY meiotic nondisjunction. Sperm disomy frequencies for chromosomes 4, 13, 18, 21 and 22 were assessed by fluorescence in situ hybridisation in four of these individuals. As a group, the Turner syndrome fathers showed a general increase in disomy frequencies for chromosomes 13, 21 and 22, with a statistically significant increase in disomy frequencies for chromosomes 13 and 22 in one of the fathers and for chromosome 21 in two of them. Data from a previous work carried out by us in two fathers of Down syndrome patients of paternal origin also revealed increased sperm disomy frequencies for chromosomes 13, 21 and 22. Pooled as one group, these six fathers of aneuploid offspring of paternal origin had a statistically significant increase in the frequency of nondisjunction for these chromosomes with respect to control individuals. Our findings indicate that there may be an association between fathering aneuploid offspring and increased frequencies of aneuploid spermatozoa. Such increases do not seem to be restricted to the chromosome pair responsible for the aneuploid offspring. Acrocentric chromosomes and other chromosome pairs that usually show only one chiasma during meiosis seem to be more susceptible to malsegregation.

Aneuploidy↗
PubMed2001

Membrane blebbing during apoptosis results from caspase-mediated activation of ROCK I.

The execution phase of apoptosis is characterized by marked changes in cell morphology that include contraction and membrane blebbing. The actin-myosin system has been proposed to be the source of contractile force that drives bleb formation, although the biochemical pathway that promotes actin-myosin contractility during apoptosis has not been identified. Here we show that the Rho effector protein ROCK I, which contributes to phosphorylation of myosin light-chains, myosin ATPase activity and coupling of actin-myosin filaments to the plasma membrane, is cleaved during apoptosis to generate a truncated active form. The activity of ROCK proteins is both necessary and sufficient for formation of membrane blebs and for re-localization of fragmented DNA into blebs and apoptotic bodies.

3T3 Cells↗
PubMed2001

Linear increase of diploidy in human sperm with age: a four-colour FISH study.

The aim of this study was to determine if donor age is associated with an increased incidence of diploidy and of disomy for the sex chromosomes and for chromosomes 6 and 21. We used simultaneous fluorescence in situ hybridisation (FISH) for chromosomes 6, 21, X and Y in sperm from 18 healthy donors, aged 24-74 years (mean 48.8 years). A total of 194 024 sperm were analysed, with a minimum of 10 000 sperm scored for each donor. Our results indicate a significant increase of the level of diploidy (P=0.002), and a marginal significance of total sex chromosome disomy (P=0.055) with age. No increase was observed for disomies XX, YY, XY, 21 or 6. The percentages of increase for disomy and for diploidy ranged from 0.3 to 17% for each 10-year period. Chromosomes 6 and 21 did not segregate preferentially with the X or Y chromosomes. Our findings show a linear trend association between age and diploidy in human males.

Adult↗
PubMed2001

Class III pistil-specific extensin-like proteins from tobacco have characteristics of arabinogalactan proteins.

Class III pistil-specific extensin-like proteins (PELPIII) are specifically localized in the intercellular matrix of tobacco (Nicotiana tabacum) styles. After pollination the majority of PELPIII are translocated into the callosic layer and the callose plugs of the pollen tubes, which could suggest a function of PELPIII in pollen tube growth. PELPIII may represent one of the chemical and/or physical factors from the female sporophytic tissue that contributes to the difference between in vivo and in vitro pollen tube growth. PELPIII glycoproteins were purified and biochemically characterized. Because of their high proline (Pro) and hydroxy-Pro (Hyp) content, PELPIII proteins belong to the class of Pro/Hyp-rich glycoproteins. The carbohydrate moiety of PELPIII is attached through O-glycosidic linkages and comprises more than one-half the total glycoprotein. Deglycosylation of PELPIII revealed two backbones, both reacting with PELPIII-specific antibodies. N-terminal amino acid sequencing of these backbones showed that PELPIII is encoded by the MG14 and MG15 genes. Two heterogeneous N-terminal sequences of MG14 and MG15, both starting downstream of the predicted signal peptide cleavage site, seem to be present, which indicates a novel N-terminal processing. Monosaccharide analysis showed that the carbohydrate moiety of PELPIII almost completely consists of arabinose and galactose in an equal molar ratio. Carbohydrate linkage analysis showed terminal and 2-linked arabinofuranosyl residues, as well as terminal and 6-, 3-, and 3,6-linked galactopyranosyl residues to be present, indicating the presence of both extensin-like and Type II arabinogalactan oligosaccharide units. The ability of beta-glucosyl Yariv reagent to bind with PELPIII confirmed the arabinogalactan protein-like characteristics of these proteins.

Amino Acid Sequence↗