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Biomedical subjects

A Ferrer

Publications and source records attributed to A Ferrer.

At least 55 records · Page 3Linked to original sources
PubMed2000

Characterization of dehydrodolichyl diphosphate synthase of Arabidopsis thaliana, a key enzyme in dolichol biosynthesis.

The enzyme dehydrodolichyl diphosphate (dedol-PP) synthase is a cis-prenyltransferase that catalyzes the synthesis of dedol-PP, the long-chain polyprenyl diphosphate used as a precursor for the synthesis of dolichyl phosphate. Here we report the cloning and characterization of a cDNA from Arabidopsis thaliana encoding dedol-PP synthase. The identity of the cloned enzyme was confirmed by functional complementation of a yeast mutant strain defective in dedol-PP synthase activity together with the detection of high levels of dedol-PP synthase activity in the transformed yeast mutant. The A. thaliana dedol-PP synthase mRNA was detected at high levels in roots but was hardly detected in flowers, leaves, stems and in A. thaliana suspension-cultured cells.

Alkyl and Aryl Transferases↗
PubMed2000

cdc25a and the splicing variant cdc25b2, but not cdc25B1, -B3 or -C, are over-expressed in aggressive human non-Hodgkin's lymphomas.

cdc25 is a family of phosphatases that activate the cyclin-dependent kinases at different points of the cell cycle. cdc25A and -B, but not -C, have been shown to have oncogenic potential. Three different splicing variants of the cdc25B gene, cdc25B1, -B2 and -B3, have also been identified. Experimental studies suggest that cdc25B2 may be more active in vivo than cdc25B3 and -B1, but the relative expression of these splicing variants in human tumors is not known. In this study, we have analyzed the expression of cdc25A, -B1, -B2, -B3 and -C mRNA in 9 non-neoplastic lymphoid samples, 89 non-Hodgki&ngrave;s lymphomas and 9 hematological cancer cell lines by semi-quantitative RT-PCR. cdc25A, -B and -C protein expression was examined by Western blot. Normal peripheral blood lymphocytes and reactive tissues expressed cdc25B1 and -B3 mRNA and very low or undetectable levels of cdc25A, -B2 and -C. High levels of cdc25A and cdc25B2 were found in 35% and 39% of the tumors, respectively, and they were more frequently observed in aggressive than in indolent lymphomas. cdc25B1 and -B3 splice variants were detected in virtually all tumors, and no significant differences were found between high- and low-grade lymphomas. cdc25A and -B protein expression was also higher in aggressive than in indolent lymphomas. cdc25C expression was relatively low in virtually all cases. In conclusion, these findings suggest that cdc25A and -B2, but not cdc25B1, -B3 and -C, are over-expressed in a relatively large number of malignant lymphomas and may participate in the pathogenesis of aggressive variants.

Blotting, Western↗
PubMed2000

[Mechanical and metabolic reproducibility of resistance test of expiratory muscles with incremental threshold loading].

BACKGROUND: The study of respiratory muscle endurance has mainly focused on inspiratory muscles. A new method to measure expiratory muscle endurance, through incremental threshold loading using a weighted plunger valve, has recently been described. OBJECTIVES: To evaluate the mechanical features of the plunger valve and the reproducibility of the method from the standpoint of both mechanics and metabolism. METHODS: Four untrained healthy subjects performed an incremental test with expiratory threshold loading (50 g every 2 min) on each of three non-consecutive days; each test continued until the subject could no longer open the valve. Mouth pressure was recorded continuously during each test; on the first two test days, oxygen consumption (VO2) was also measured. RESULTS: Opening and closing pressures were the same and were independent of expiratory flow, with a linear load-pressure relationship (4 cmH2O) for every 10 g of weight). The maximal tolerated load (MTL) in the three tests was stable for two of the subjects, whereas the maximal load was reached by the other two subjects in the second and third tests, respectively. When MTL was reached in the third test, mean and peak mouth pressures (the latter expressed as percent of maximal expiratory pressure [MEP]) were 49 +/- 4% and 71 +/- 4%, respectively; the expiratory tension-time index measured at the mouth ([PMEANmouth/MEP] x [TE/Ttot]) was 0.25 +/- 0.02 (TE: expiratory time; Ttot: total time). In the first and second tests, we also measured oxygen consumption of the recruited muscles, which were mainly the expiratory muscles (VO2respmax); consumption in the last test was 213 +/- 65 ml O2/min (2.9 +/- 1.1 ml O2/kg/min). The intraindividual coefficient of variation ranged from 6.3% to 19.5% for the mechanical parameters and from 14% to 21% for the metabolic ones. CONCLUSIONS: The expiratory endurance test using a threshold valve allows quantification of muscle and metabolic reserve under incremental expiratory loads. The valve has appropriate mechanical characteristics for this purpose and reproducibility is acceptable, through the precise determination of the may require up to three tests.

Adult↗
PubMed2000

Molecular cloning and expression analysis of the mevalonate kinase gene from Arabidopsis thaliana.

Mevalonate kinase (MVK), the enzyme that catalyzes the phosphorylation of mevalonate to produce mevalonate 5-phosphate, is considered as a potential regulatory enzyme of the isoprenoid biosynthetic pathway. The Arabidopsis thaliana MVK gene corresponding to the MVK cDNA previously isolated has been cloned and characterized. RNAse protection analysis indicated that the expression of the MVK gene generates three mRNA populations with 5' ends mapping 203, 254 and 355 nt upstream of the MVK ATG start codon. Northern blot analysis showed that the MVK mRNA accumulates preferentially in roots and influorescences. Histochemical analysis, with transgenic A. thaliana plants containing a translational fusion of a 1.8 kb fragment of the 5' region of the MVK gene to the beta-glucuronidase (GUS) reporter gene, indicated that the MVK 5'-flanking region directs widespread expression of the GUS gene throughout development, although the highest levels of GUS activity are detected in roots (meristematic region) and flowers (sepals, petals, anthers, style and stigmatic papillae). The expression pattern of the MVK gene suggests that the role of the encoded MVK is the production of a general pool of mevalonate-5-phosphate for the synthesis of different classes of isoprenoids involved in both basic and specialized plant cell functions. Functional promoter deletion analysis in transfected A. thaliana protoplasts indicated that regulatory elements between positions -295 and -194 of the MVK 5'-flanking region are crucial for high-level MVK gene expression.

Amino Acid Sequence↗
PubMed2000

The Arabidopsis thaliana PPX/PP4 phosphatases: molecular cloning and structural organization of the genes and immunolocalization of the proteins to plastids.

The PPX/PP4 Ser/Thr protein phosphatases belong to the type 2A phosphatase subfamily and are present in most eukaryotic organisms. We have previously isolated two closely related DNAs encoding PPX isoforms (PPX-1 and PPX-2) of Arabidopsis thaliana. Here we report the molecular cloning of the genes encoding these proteins. The genes PPX-1 and PPX-2 are composed of eight exons and seven introns located at equivalent positions related to the coding sequences. Whereas the intron-exon organization of the PPX genes is completely different from that of the PP2A-3/PP2A-4 A. thaliana family, specific intron-exon boundaries are conserved among PPX genes from distantly related organisms. Based on GUS expression, both PPX genes show the same spatial and temporal pattern of expression: they are expressed in all the organs and tissues analyzed, and from the earliest stage of development. When PPX proteins were localized to the root in semi-thin methacrylate sections by immunofluorescence, staining was predominantly confined to small organelles, shown to be plastids by co-localization of PPX and ferredoxin. Interestingly, only some ferredoxin-positive plastids were also PPX-positive, and PPX staining was consistently brighter in the epidermis. The localization was confirmed with immunogold and electron microscopy. Our results suggest that, despite its strong sequence conservation, PPX in plants functions differently than in animals.

Arabidopsis↗
PubMed2000

Spatial and temporal patterns of GUS expression directed by 5' regions of the Arabidopsis thaliana farnesyl diphosphate synthase genes FPS1 and FPS2.

Farnesyl diphosphate synthase (FPS), the enzyme that catalyses the synthesis of farnesyl diphosphate (FPP) from isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP), is considered a regulatory enzyme of plant isoprenoid biosynthesis. The promoter regions of the FPS1 and FPS2 genes controlling the expression of isoforms FPS1S and FPS2, respectively, were fused to the beta-glucuronidase (GUS) reporter gene and introduced into Arabidopsis thaliana plants. The FPS1S:GUS gene is widely expressed in all plant tissues throughout development, thus supporting a role for FPS1S in the synthesis of isoprenoids serving basic plant cell functions. In contrast, the FPS2:GUS gene shows a pattern of expression restricted to specific organs at particular stages of development. The highest levels of GUS activity are detected in flowers, especially in pollen grains, from the early stages of flower development. After pollination, much lower levels of GUS activity are detected in the rest of floral organs, with the exception of the ovary valves, which remain unstained throughout flower development. GUS activity is also detected in developing and mature seeds. In roots, GUS expression is primarily detected at sites of lateral root initiation and in junctions between primary and secondary roots. No GUS activity is detected in root apical meristems. GUS expression is also observed in junctions between primary and secondary stems. Overall, the pattern of expression of FPS2:GUS suggests a role for FPS2 in the synthesis of particular isoprenoids with specialized functions. Functional FPS2 gene promoter deletion analysis in transfected protoplasts and transgenic A. thaliana plants indicate that all the cis-acting elements required to establish the full pattern of expression of the FPS2 gene are contained in a short region extending from positions -111 to +65. The potential regulatory role of specific sequences within this region is discussed.

Alkyl and Aryl Transferases↗
PubMed2000

Immune responses to dystropin: implications for gene therapy of Duchenne muscular dystrophy.

Introduction of dystrophin by gene transfer into the dystrophic muscles of Duchenne muscular dystrophy (DMD) patients has the possibility of triggering an immune response as many patients will not have been exposed to some (or all) of the epitopes of dystrophin. This could in turn lead to cytotoxic destruction of transfected muscle fibres. We assessed such concerns in the dystrophin-deficient mdx mouse using plasmid DNA as the gene transfer system. This avoids complications associated with the administration of viral proteins. Gene transfer of cDNAs encoding mouse full-length or a truncated minidystrophin did not evoke either a humoral or cytotoxic immune response. Mdx mice may be tolerant due to the presence of rare 'revertant' dystrophin-positive fibres in their skeletal muscles. In contrast, gene transfer of human full-length or minidystrophin provoked both humoral and cytotoxic responses leading to destruction of the transfected fibres. These experiments demonstrate the potential risk of deleterious effects following gene therapy in DMD patients and lead us to suggest that patients enrolled in gene therapy trials should ideally have small, preferably point, mutations and evidence of 'revertant' dystrophin-positive muscle fibres.

Analysis of Variance↗
PubMed2000

Isoenzymes of human lice: pediculus humanus and P. capitis.

Human lice (Phthiraptera: Pediculidae) from Africa, America and Europe were electrophoresed for 28 enzymes, with special interest in metabolic factors likely to be involved with insecticide resistance. Zymogram profiles of the body louse (Pediculus humanus L. from France and U.S.A.) and the head louse (P. capitis DeGeer from France, Madagascar, Mali & Senegal) were compared. Only esterase two enzymes, phosphoglucomutase (Pgm) and 3 (Est-3), showed electrophoretic variation. In our starch gel electrophoresis conditions, P. humanus showed three electromorphs of Pgm migrating anodally 6, 11 and 16 mm (designated alleles a, b, c, respectively). Of the putative Pgm alleles, b and c occurred in all samples of both species of lice, whereas allele a was found only in P. humanus lab strain from U.S.A. Esterase 3 had four electromorphs migrating 23, 26, 30 and 35 mm (designated alleles a, b, c and d). Among putative Est alleles, a was found only in P. capitis from Bamako (all 14 specimens aa homozygotes), allele d was found only in P. capitis from Dakar (39% frequency), whereas Est-3 alleles b and c showed apparently balanced polymorphism in all samples of both P. humanus and P. capitis except that from Bamako. Despite the limited amount of isoenzyme variation detected (only 2/31 polymorphic loci), divergences of Est-3 and Pgm among Pediculus populations may be relevant to their biosystematics and resistance.

Animals↗
PubMed2000

Nosocomial Legionnaire's disease in a children's hospital.

BACKGROUND: Only a few cases of nosocomial Legionella sp. infection have been reported in children. We report the clinical and epidemiologic data of five nosocomial legionellosis cases that occurred in the Pediatric Nephrology Service between August, 1994, and December, 1998, and the control measures adopted. METHODS: The Hospital Materno-Infantil Vall d'Hebron, Barcelona, is a 407-bed tertiary care hospital. The pediatric kidney transplant unit has three isolated beds in the same ward within the Pediatric Nephrology Service. Diagnostic workup to establish Legionella pneumophila infection included culture, fluorescent antibody and serologic studies. Macrorestriction analysis of genomic DNA was used as epidemiologic markers of the isolated strains. RESULTS: In May, 1996, a case of L. pneumophila serogroup 6 pneumonia was identified in a 19-year-old youth who had received a kidney transplant 16 days earlier. Retrospective and prospective analysis of legionellosis cases diagnosed at our center up to August, 1994, yielded four additional cases. Four patients had had a kidney transplant and were receiving immunosuppressive therapy, and the fifth had been diagnosed with systemic lupus erythematosus with renal involvement. L. pneumophila serogroup 6 was isolated in bronchial secretions in four cases; in the fifth patient the diagnosis was made by serology. L. pneumophila serogroup 6 was isolated from potable water of the hospital. Molecular epidemiologic methods revealed the identity of the environmental and clinical isolates. Showering was implicated as the most feasible means of exposure to contaminated water. CONCLUSIONS: Nosocomial legionellosis, albeit rare in children, should be considered in the differential diagnosis of pneumonias, particularly in immunosuppressed children, because the fatality rate may be high without early diagnosis and treatment.

Adolescent↗
PubMed2000

Detection of Pseudomonas savastanoi pv. savastanoi in olive plants by enrichment and PCR.

The sequence of the gene iaaL of Pseudomonas savastanoi EW2009 was used to design primers for PCR amplification. The iaaL-derived primers directed the amplification of a 454-bp fragment from genomic DNA isolated from 70 strains of P. savastanoi, whereas genomic DNA from 93 non-P. savastanoi isolates did not yield this amplified product. A previous bacterial enrichment in the semiselective liquid medium PVF-1 improved the PCR sensitivity level, allowing detection of 10 to 100 CFU/ml of plant extract. P. savastanoi was detected by the developed enrichment-PCR method in knots from different varieties of inoculated and naturally infected olive trees. Moreover, P. savastanoi was detected in symptomless stem tissues from naturally infected olive plants. This enrichment-PCR method is more sensitive and less cumbersome than the conventional isolation methods for detection of P. savastanoi.

Bacterial Proteins↗
PubMed2000

Optimizing ammonia pressurization/depressurization processing conditions to enhance enzymatic susceptibility of dwarf elephant grass.

An ammonia pressurization/depressurization process was investigated to evaluate the potential of producing reducing sugars from dwarf elephant grass, a warm-season forage. Moisture, temperature, and ammonia loading affected sugar yield (p < 0.0001). At optimal conditions, ammonia processing solubilized 50.9% of the hemicellulose and raised the sugar yield (percentage of theoretical) from 18 to 83%. Glucose and xylose production were increased 3.2- and 8.2-fold, respectively. The mild processing conditions of the ammonia treatment (90-100 degrees C, 5 min), the low enzyme loading (2 international filter paper units/g), and the short hydrolysis time (24 h), greatly enhance the potential of using forages to produce sugars valuable for several applications.

Ammonia↗
PubMed2000

Early puberty: rapid progression and reduced final height in girls with low birth weight.

OBJECTIVE: To assess whether, in girls with early onset of puberty, low birth weight is a risk factor for rapid progression to menarche and for short adult stature. DESIGN: Longitudinal clinical assessment of 54 Catalan (Northern Spanish) girls followed from early onset of puberty (onset of breast development between 8.0 and 9.0 years of age) to final height. The timing of menarche and the final height were analyzed a posteriori according to birth weight, the cutoff level between normal and low birth weight subgroups being -1.5 standard deviation (SD; approximately 2.7 kg at term birth). RESULTS: Normal and low birth weight girls had similar target heights and characteristics at diagnosis of early puberty. However, menarche occurred on average 1.6 years earlier in low versus normal birth weight girls (11.3 +/-.3 years vs 12.9 +/-.2 years), and final height was >5 cm shorter in low birth weight girls (parental adjusted height SD: -.6 +/-.2 cm vs.3 +/-.2 cm). CONCLUSION: The timing of menarche and the level of final height in Catalan girls with early onset of puberty was found to depend on prenatal growth. Girls with normal birth weight tend to progress slowly through puberty with a normal timing of menarche and normal final height. In contrast, girls with low birth weight tend to progress relatively rapidly to an early menarche and to a reduced final height. If these findings are confirmed in other ethnic and/or larger groups, then a subgroup has been identified that will most likely benefit from any therapeutic intervention aiming at a delay of pubertal development and/or an increase of final height.

Body Height↗
PubMed2000

Intervention study for smoking cessation in diabetic patients: a randomized controlled trial in both clinical and primary care settings.

OBJECTIVE: To evaluate the effectiveness of a nurse-managed smoking cessation intervention in diabetic patients. RESEARCH DESIGN AND METHODS: This randomized controlled clinical trial involved 280 diabetic smokers (age range 17-84 years) who were randomized either into control (n = 133) or intervention (n = 147) groups at 12 primary care centers and 2 hospitals located in Navarre, Spain. The intervention consisted of a 40-min nurse visit that included counseling, education, and contracting information (a negotiated cessation date). The follow-up consisted of telephone calls, letters, and visits. The control group received the usual care for diabetic smokers. Baseline and 6-month follow-up measurements included smoking status (self-reported cessation was verified by urine cotinine concentrations), mean number of cigarettes smoked per day, and stage of change. RESULTS: At the 6-month follow-up, the smoking cessation incidence was 17.0% in the intervention group compared with 2.3% in the usual care group, which was a 14.7% difference (95% CI 8.2-21.3%). Among participants who continued smoking, a significant reduction was evident in the average cigarette consumption at the 6-month follow-up. The mean number of cigarettes per day decreased from 20.0 at baseline to 15.5 at 6 months for the experimental group versus from 19.7 to 18.1 for the control group (P < 0.01). CONCLUSIONS: A structured intervention managed by a single nurse was shown to be effective in changing the smoking behavior of diabetic patients.

Adolescent↗
PubMed2000

[Microbiological study of post mortem lung biopsy in pediatric patients].

AIM: The aim of this study was to correlate the morphologic and microbiologic findings of the post mortem biopsies of a series of pediatric patients. MATERIAL AND METHODS: A complete morphologic study of all the organs of the patients included in the study was performed. In the pulmonary samples provided by the pathologist, the presence of aerobic and anaerobic microorganisms and fungi were determined. RESULTS: Ninety-three pulmonary biopsies corresponding to 77 dead patients (47 infants and 30 fetuses) were undertaken. Forty-five patients showed pulmonary histology of infectious processes. The concordance between the morphology and the microbiologic culture was of 66.2%. The microorganisms most frequently isolated were gram positive cocci (46.4%) followed by gram negative bacillus (30.4%). CONCLUSIONS: Although post mortem pulmonary tissue cultures should be interpreted with caution without first relating these with the morphologic findings and the clinical characteristics of the patient, a statistically significant agreement was observed between the anatomo-pathological study and the microbiologic findings.

Adolescent↗
PubMed2000

[Intermittent pre-excitation after adenosine administration].

We describe a case of 2:1 intermittent preexcitation after adenosine administration in a patient with an accessory pathway that did not show preexcitation on the basal ECG. We review the mechanisms involved that explain this event and the possible utility of adenosine to show accessory pathways that do not show preexcitation on the ECG.

Adenosine↗
PubMed2000

[Microorganisms isolated in cases of pertussis-like syndrome].

OBJECTIVE: To describe the etiologic study of the pertussis-like syndrome, not only as far as Bordetella genus is concerned but also regarding the causative role of other microorganisms for a 11-year period (1988-1998). METHODS: In all specimens from patients suffering from pertussis-like cough the presence of Bordetella spp., other bacteria, viruses, and mycoplasma was investigated. The analysed data included microbiological findings and epidemiologic issues (age, sex, hospital admission area, yearly distribution and seasonal period). RESULTS: A total of 1,063 specimens were investigated, most of them nasopharyngeal aspirates (910), corresponding to 905 patients; a positive culture was obtained form 56.9 of these patients. B. pertussis was isolated from 10.5% of patients. As for other bacteria, Haemophilus influenzae and Streptococcus pneumoniae were also isolated, in 16.9% and 15.8% of occasions, respectively. The respiratory syncitial virus was isolated from 10.7% of patients and other viruses in 9.4%. Among mycoplasma, Ureaplasma urealyticum predominated, with a recovery rate of 2.9%. The male/female ratio was 495/410; the ages of 67.2% of patients ranged from 0 to 6 months; a total of 689 (76.1%) required hospital admission. The recovery of B. pertussis and adenoviruses predominated during spring and summer months. In contrast, H. influenzae, S. pneumoniae and respiratory syncitial virus were recovered more frequently during winter months. CONCLUSIONS: Most patients with pertussis-like syndrome are children aged less than 6 months. The recovery percentages of B. pertussis and respiratory syncitial virus are identical and therefore we think that the investigation of their presence in this syndrome is fully warranted as well as the search for other microorganisms, since clinical symptoms are commonly non-specific among infants.

Age Distribution↗