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Biomedical subjects

E Carrera

Publications and source records attributed to E Carrera.

At least 19 recordsLinked to original sources

Continuous assessment of electrical epileptic activity in acute stroke.

OBJECTIVE: To determine the incidence and risk factors of electrical seizures and other electrical epileptic activity using continuous EEG (cEEG) in patients with acute stroke. METHODS: One hundred consecutive patients with acute stroke admitted to our stroke unit underwent cEEG using 10 electrodes. In addition to electrical seizures, repetitive focal sharp waves (RSHWs), repetitive focal spikes (RSPs), and periodic lateralized epileptic discharges (PLEDs) were recorded. RESULTS: In the 100 patients, cEEG was recorded for a mean duration of 17 hours 34 minutes (range 1 hour 12 minutes to 37 hours 10 minutes). Epileptic activity occurred in 17 patients and consisted of RSHWs in seven, RSPs in seven, and PLEDs in three. Electrical seizures occurred in two patients. On univariate Cox regression analysis, predictors for electrical epileptic activity were stroke severity (high score on the National Institutes of Health Stroke Scale) (hazard ratio [HR] 1.12; p = 0.002), cortical involvement (HR 5.71; p = 0.021), and thrombolysis (HR 3.27; p = 0.040). Age, sex, stroke type, use of EEG-modifying medication, and cardiovascular risk factors were not predictors of electrical epileptic activity. On multivariate analysis, stroke severity was the only independent predictor (HR 1.09; p = 0.016). CONCLUSION: In patients with acute stroke, electrical epileptic activity occurs more frequently than previously suspected.

Adult↗

[Neurology].

In this paper, we present the last studies with impact on daily practice and up-to-date guidelines which can lead to a rational approach of the patient with neurological disease. In cerebrovascular diseases, several strategies are discussed to increase the number of patients who can benefit from thrombolysis, whereas in secondary prevention, the association of clopidogrel and aspirin versus clopidogrel has not shown to be beneficial. In Parkinson's disease, the inclusion criteria and benefits of deep brain stimulation are more precisely known. New genetic targets have been found in the field of myopathies, which open new therapeutic perspectives.

Brain Diseases↗

Adherence to treatment in inflammatory bowel disease.

AIM: Adherence to therapy is important to ensure success. We wanted to explore this feature in patients with inflammatory bowel disease. PATIENTS AND METHODS: We explored adherence to treatment and its modifiers in 40 patients with inflammatory bowel disease using a battery of tests. RESULTS: A 67% of patients (95% CI: 51-81%) acknowledged a certain degree of involuntary nonadherence, and 35% (95% CI: 20-51%) of voluntary nonadherence. Overall, 72% (95% CI: 56-85%) of patients had some form of nonadherence. An objective correlation of these self-reported data was assessed by the determination of urine salicylate levels in the subset of patients treated with mesalazine or its derivatives (15 cases). Two of them (13%) had no detectable urinary drug levels, indicating complete nonadherence. Voluntary nonadherence was higher in patients with lower scores in the intestinal (p = 0.02) and social areas (p = 0.015) of IBDQ-32, as well as in those with less active Crohn s disease (p < 0.005), patients with high depression scores and high patient-physician discordance (p = 0.01), patients with long-standing disease (p = 0.057), patients who considered themselves not to be well informed about the treatment they were getting (p = 0.04) or who trusted their attending physicians less (p = 0.03). CONCLUSIONS: Intentional nonadherence to therapy is prevalent among patients with inflammatory bowel disease. A correction of factors associated to poor adherence could lead to higher therapeutic success.

Adult↗

Efficacy and safety of thiopurinic immunomodulators (azathioprine and mercaptopurine) in steroid-dependent ulcerative colitis.

BACKGROUND: The efficacy of azathioprine in the management of steroid-dependent ulcerative colitis is taken for granted. However, study populations frequently include together steroid-dependent and refractory patients. AIM: To assess the efficacy and safety of thiopurinic immunomodulators in strictly defined steroid-dependent ulcerative colitis. METHODS: Survey of 34 patients with steroid-dependent ulcerative colitis, treated with azathioprine according to protocol. Therapeutical success: glucocorticoid withdrawal within 12 months, without steroid requirements during another year. RESULTS: Mean age was 39.1 +/- 17 years. Pancolitis and extensive colitis accounted for 50% of cases. Therapeutic success of immunomodulator treatment reached 70.6%, intention to treat analysis (confidence interval 95%: 52-84%) and 72.7%, as per protocol (confidence interval 95%: 54-86%). Mean time to steroid withdrawal was 4.6 months. In therapy successes, mean corpuscular volume and total serum bilirubin increased with treatment time (P = 0.0001). Fifteen adverse effects were observed in 13 patients (38%). Azathioprine was withdrawn in seven cases (20.6%); in four of them (with liver toxicity), treatment with mercaptopurine was indicated. CONCLUSIONS: Therapy with thiopurinic immunomodulators (azathioprine) represents the first option in the management of steroid-dependent ulcerative colitis. Its efficacy (70%) and its acceptable safety support this view. Increasing mean corpuscular volume and serum bilirubin values may be a surrogate marker of a beneficial effect.

Adult↗

Pyoderma gangrenosum associated with ulcerative colitis: response to infliximab.

Pyoderma gangrenosum is an extraintestinal manifestation of inflammatory bowel disease that can be therapeutically troublesome. We comment on the case of a patient with clinically inactive ulcerative colitis who progressively developed necrotic lesions on both tibial aspects of his legs, which corresponded both clinically and histologically to pyoderma gangrenosum. Treatment with steroids and azathioprine could not control this complication. A single dose of infliximab 5 mg/kg was given, achieving an impressive response of the skin lesions followed by complete healing 3 months later. Infliximab can be useful in the management of refractory extraintestinal manifestations of inflammatory bowel disease.

Antibodies, Monoclonal↗

The P450-4 gene of Gibberella fujikuroi encodes ent-kaurene oxidase in the gibberellin biosynthesis pathway.

At least five genes of the gibberellin (GA) biosynthesis pathway are clustered on chromosome 4 of Gibberella fujikuroi; these genes encode the bifunctional ent-copalyl diphosphate synthase/ent-kaurene synthase, a GA-specific geranylgeranyl diphosphate synthase, and three cytochrome P450 monooxygenases. We now describe a fourth cytochrome P450 monooxygenase gene (P450-4). Gas chromatography-mass spectrometry analysis of extracts of mycelia and culture fluid of a P450-4 knockout mutant identified ent-kaurene as the only intermediate of the GA pathway. Incubations with radiolabeled precursors showed that the metabolism of ent-kaurene, ent-kaurenol, and ent-kaurenal was blocked in the transformants, whereas ent-kaurenoic acid was metabolized efficiently to GA(4). The GA-deficient mutant strain SG139, which lacks the 30-kb GA biosynthesis gene cluster, converted ent-kaurene to ent-kaurenoic acid after transformation with P450-4. The B1-41a mutant, described as blocked between ent-kaurenal and ent-kaurenoic acid, was fully complemented by P450-4. There is a single nucleotide difference between the sequence of the B1-41a and wild-type P450-4 alleles at the 3' consensus sequence of intron 2 in the mutant, resulting in reduced levels of active protein due to a splicing defect in the mutant. These data suggest that P450-4 encodes a multifunctional ent-kaurene oxidase catalyzing all three oxidation steps between ent-kaurene and ent-kaurenoic acid.

Amino Acid Sequence↗

Changes in GA 20-oxidase gene expression strongly affect stem length, tuber induction and tuber yield of potato plants.

Gene StGA20ox1 encoding potato GA 20-oxidase is expressed to relatively high levels in leaves and regulated by daylength. To investigate whether this gene is involved in photoperiodic regulation of tuber formation, we have obtained transgenic potato plants expressing sense and antisense copies of the StGA20ox1 cDNA. Over-expression of this cDNA resulted in taller plants that required a longer duration of a short day photoperiod (SD) to tuberize. Tubers from these plants had a decreased time of dormancy and developed sprouts with elongated internodes. Plants expressing antisense copies of the StGA20ox1 cDNA had shorter stems, a decreased length of the internodes and tuberized earlier than control plants, showing increased tuber yields. Antisense inhibition of this gene had no visible effect on the time of dormancy of the tubers, although at the end of dormancy these formed sprouts with shortened internodes. Decreased levels of endogenous GA20 and GA1 were detected in the apex and first leaves of the antisense lines. These results demonstrate the involvement of the GA 20-oxidase activity encoded by StGA20ox1 in the control of stem elongation and in tuber induction but not in tuber dormancy, indicating that the latter may be regulated by another member of the gene family.

DNA, Complementary↗

Regulation of transcript levels of a potato gibberellin 20-oxidase gene by light and phytochrome B.

Up to three gibberellin (GA) 20-oxidase genes have now been cloned from several species including Arabidopsis, bean (Phaseolus vulgaris), and potato (Solanum tuberosum). In each case the GA 20-oxidase genes exhibit different patterns of tissue expression. We have performed extensive northern analysis on one of the potato GA 20-oxidase genes (StGA20ox1), which is the only one that shows significant transcript levels in leaves. We show that levels of StGA20ox1 transcript are elevated in transgenic antisense plants that have reduced levels of phytochrome B (PHYB) compared with wild-type plants, implicating PHYB in the control of GA biosynthesis. We show that StGA20ox1 transcript levels vary in leaves of different age throughout the plant and cycle throughout the day, furthermore they are up-regulated by light and down-regulated in the dark. The degree of the response to the light-on signal is similar in potato plants deficient in phytochrome A or PHYB and wild-type plants. The induction of StGA20ox1 by blue light raises the possibility that a blue light receptor may be involved in the control of this gene by light.

Antisense Elements (Genetics)↗

Identification of sole (Solea solea) and Greenland halibut (Reinhardtius hippoglossoides) by PCR amplification of the 5S rDNA gene.

Polymerase chain reaction (PCR) amplification of the nuclear 5S rDNA gene, has been used for the identification of sole (Solea solea) and Greenland halibut (Reinhardtius hippoglossoides). Two species-specific primers were designed to amplify specific fragments of the 5S rDNA gene in each species. The remarkably different size of the amplicons obtained gives, by simple agarose gel electrophoresis, two distinguishable band patterns for both flatfish species. This genetic marker can be very useful for the accurate identification of S. solea and Greenland halibut, to enforce labeling regulations.

Animals↗

Rat liver contains age-regulated cytosolic 3-deoxy-D-glycero-D-galacto-non-2-ulopyranosonic acid (Kdn).

Kdn (3-deoxy-D-glycero-D-galacto-non-2-ulopyranosonic acid), a unique deaminated member of the sialic acid family, has emerged as a new building block of glycoconjugates from a wide variety of organisms, ranging from bacteria to mammals. In particular, the presence of Kdn has been demonstrated in different rat organs and tissues, but not in liver. Here we report on the detection and quantitation of Kdn in rat liver and on its variations with postnatal development and aging. We have previously established the optimal conditions for derivatization of Kdn with 1,2-diamino-4, 5-methylene-dioxybenzene (DMB), and detection by reverse-phase HPLC. Analysis of whole liver homogenates and different subcellular fractions reveals that Kdn is fundamentally present in the cytosolic fraction as nucleotide precursor. The expression of Kdn, Neu5Gc, and Neu5Ac changes unevenly with age. While the content of Neu5Ac, the major species, and Neu5Gc decreases to a different extent from newborn to old animals, Kdn content decreases from newborn to trace amounts in adult rats and increases again with aging. Thus, expression of Kdn, Neu5Gc, and Neu5Ac appears to be independently regulated.

Aging↗

Feedback control and diurnal regulation of gibberellin 20-oxidase transcript levels in potato.

Tuber formation in potato (Solanum tuberosum) is promoted by short photoperiods and is inhibited by gibberellins (GAs). Endogenous levels of GA1 were shown to decrease in stolons and leaves of potato plants induced to tuberize, which suggests that photoperiodic regulation of GA biosynthesis may play a role in tuber induction. We report the isolation of three potato cDNA clones (StGA20ox1-3) encoding GA 20-oxidase, a key regulatory enzyme in the GA-biosynthetic pathway. Using northern analysis, we detected a differential pattern of tissue-specific expression of the mRNAs corresponding to these clones. StGA20ox mRNAs were also very abundant in leaves of the potato ga1 mutant, which is blocked in the 13-hydroxylation step, and were strongly down-regulated by gibberellic acid, suggesting a feedback regulation of these genes. In plants grown in short-day (inductive) conditions, levels of the StGA20ox transcripts in leaves fluctuated during a 24-h period, with a peak of accumulation observed about 4 h after the lights were turned off. Interruption of the night with a 30-min "night break" of light (noninductive conditions) did not have a marked effect on the levels of accumulation of the three GA 20-oxidase mRNAs during the day, but it induced a second peak of expression of StGA20ox1 and StGA20ox3 transcripts late in the night. This observation, together with the finding that StGA20ox1 mRNA is expressed at high levels in leaves, suggests that night-break induction of this gene might play a role in the control of tuberization by regulating endogenous levels of GAs in response to daylength conditions.

Amino Acid Sequence↗

Indirect enzyme-linked immunosorbent assay for the identification of sole (Solea solea), European plaice (Pleuronectes platessa), flounder (Platichthys flesus), and Greenland halibut (Reinhardtius hippoglossoides).

Polyclonal antibodies produced against soluble muscle protein extracts from sole (Solea solea), European plaice (Pleuronectes platessa), flounder (Platichthys flesus), and Greenland halibut (Reinhardtius hippoglossoides) were used in an indirect enzyme-linked immunosorbent assay for the specific identification of fillets from these flatfish species. The assay was performed in two different formats: microtiter plates and immunostick tubes. Immunorecognition of antibodies adsorbed to their specific fish samples was made with goat antirabbit immunoglobulins conjugated to the enzyme horseradish peroxidase. Subsequent enzymatic conversion of the substrate allowed unequivocal identification of all flatfish species studied.

Animals↗

Antimicrobial susceptibility of anaerobic and aerobic bacteria isolated from patients with mixed infections in Nicaragua.

The agar dilution method was used to test the activity of ampicillin, benzylpenicillin, cefoxitin, imipenem, clindamycin, metronidazole, chloramphenicol, gentamicin, methicillin and vancomycin against 241 anaerobic and 227 aerobic bacteria isolated from 136 patients with intraabdominal infections and 49 with nonintraabdominal infections. Beta-lactamase production was tested in all strains. Overall, imipenem, metronidazole and chloramphenicol were the most active antimicrobial agents against anaerobic bacteria followed by clindamycin. Only the Bacteroides fragilis group was shown to be less susceptible to clindamycin (MIC90 8 mg/l). Ampicillin and cefoxitin were the least active beta-lactam antibiotics against the most common isolated B. fragilis group strains (MIC(90) >1024 and 64 mg/l, respectively) and against Escherichia coli strains (MIC(90) >1024 and >1024 mg/l, respectively). Chloramphenicol showed low activity against the Gram-negative aerobic bacteria, while gentamicin had good activity against the aerobic bacteria tested, except for E. coli and Pseudomonas. Among the Gram-positive aerobic and anaerobic bacteria tested, Staphylococcus aureus was shown to be less susceptible to beta-lactam antibiotics (29% were methicillin resistant). No vancomycin-resistant S. aureus strains were found. A good correlation between beta-lactamase production and beta-lactam resistance was observed.

Bacteria, Aerobic↗

Antimicrobial susceptibility of anaerobicand aerobic bacteria isolated from patientswith mixed infections in Nicaragua.

The agar dilution method was used to test the activity of ampicillin, benzylpenicillin, cefoxitin, imipenem, clindamycin, metronidazole, chloramphenicol, gentamicin, methicillin and vancomycin against 241 anaerobic and 227 aerobic bacteria isolated from 136 patients with intraabdominal infections and 49 with nonintraabdominal infections. Beta-lactamase production was tested in all strains. Overall, imipenem, metronidazole and chloramphenicol were the most active antimicrobial agents against anaerobic bacteria followed by clindamycin. Only the Bacteroides fragilis group was shown to be less susceptible to clindamycin (MIC90 8 mg/l). Ampicillin and cefoxitin were the least active beta-lactam antibiotics against the most common isolated B. fragilis group strains (MIC(90) >1024 and 64 mg/l, respectively) and against Escherichia coli strains (MIC(90) >1024 and >1024 mg/l, respectively). Chloramphenicol showed low activity against the Gram-negative aerobic bacteria, while gentamicin had good activity against the aerobic bacteria tested, except for E. coli and Pseudomonas. Among the Gram-positive aerobic and anaerobic bacteria tested, Staphylococcus aureus was shown to be less susceptible to beta-lactam antibiotics (29% were methicillin resistant). No vancomycin-resistant S. aureus strains were found. A good correlation between beta-lactamase production and beta-lactam resistance was observed.

Journal Article↗

Expression of the Arabidopsis abi1-1 mutant allele inhibits proteinase inhibitor wound-induction in tomato.

Abscisic acid (ABA) is an essential component in the wound signalling cascade. Increased levels of endogenous ABA were observed after wounding and shown to be a requisite for wound-induced expression of the proteinase inhibitor II genes. We have taken advantage of the dominant character of the Arabidopsis abi1-1 mutation, to investigate whether ABl1 has a function in ABA signalling in response to wounding. Transgenic tomato plants carrying copies of either the wild-type ABI1 or the mutant abi1-1 alleles were obtained and assayed for wound-induction of the pin2 or LAP genes. While normal levels of gene induction were observed in the transgenic ABI1 plants, the abi1-1 transformants displayed a severe wilty phenotype and reduced seed dormancy. Expression of the abi1-1 dominant mutation blocked accumulation of the drought-induced TAS14 and LE25 mRNAs in response to ABA, as well as ABA- and wound-induced expression of the defense-associated pin2 and LAP transcripts. MeJA-induction of the pin2 and LAP mRNAs, on the contrary, was not affected in the abi1-1 transformants. These results indicate that abi1-1 inhibits wound-induced expression of the pin2 and LAP transcripts by blocking ABA-induction of these genes. This implicates ABI1 in wound-signalling and suggests that a common early ABA signalling pathway may function in the responses to wounding and water stress.

Alleles↗

Polymerase chain reaction-restriction fragment length polymorphism analysis of a short fragment of the cytochrome b gene for identification of flatfish species.

Restriction site analysis of polymerase chain reaction (PCR) products from a conserved region of the cytochrome b gene has been used for the specific identification of sole (Solea solea), European plaice (Pleuronectes platessa), flounder (Platichthys flesus), and Greenland halibut (Reinhardtius hippoglossoides). PCR amplification of the cytochrome b gene using a universal primer together with a primer specifically designed as a part of this study produced a 201-bp fragment in all species analyzed. Digestions of the PCR products with Sau3Al, BsmAl, Rsal, and Mn/l endonucleases, followed by agarose gel electrophoresis of the digested PCR products, yielded specific profiles that enabled direct identification of each species analyzed.

Animals↗

Identification of Atlantic salmon (Salmo salar) and rainbow trout (Oncorhynchus mykiss) by using polymerase chain reaction amplification and restriction analysis of the mitochondrial cytochrome b gene.

Restriction site analysis of polymerase chain reaction (PCR) products from a conserved region of the cytochrome b gene has been used for the identification of fresh and smoked samples of Atlantic salmon (Salmo salar) and rainbow trout (Oncorhynchus mykiss). Digestion of the 359-bp PCR product with the endonucleases EcoRV and TaqI yielded specific banding patterns for salmon and trout. This genetic marker can be very useful for detecting fraudulent substitution of the cheaper smoked trout for the more expensive smoked salmon.

Animals↗