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Biomedical subjects

J L Azevedo

Publications and source records attributed to J L Azevedo.

At least 19 recordsLinked to original sources
PubMed2005

Rapid, specific and quantitative assays for the detection of the endophytic bacterium Methylobacterium mesophilicum in plants.

Xylella fastidiosa is a xylem-limited bacterium that causes citrus variegated chlorosis disease in sweet orange. There is evidence that X. fastidiosa interacts with endophytic bacteria present in the xylem of sweet orange, and that these interactions, particularly with Methylobacterium mesophilicum, may affect disease progress. However, these interactions cannot be evaluated in detail until efficient methods for detection and enumeration of these bacteria in planta are developed. We have previously developed standard and quantitative PCR-based assays specific for X. fastidiosa using the LightCycler system [Li, W.B., Pria Jr., L.P.M.W.D., X. Qin, and J.S. Hartung, 2003. Presence of Xylella fastidiosa in sweet orange fruit and seeds and its transmission to seedlings. Phytopathology 93:953-958.], and now report the development of both standard and quantitative PCR assays for M. mesophilicum. The assays are specific for M. mesophilicum and do not amplify DNA from other species of Methylobacterium or other bacteria commonly associated with citrus or plant tissue. Other bacteria tested included Curtobacterium flaccumfaciens, Pantoea agglomerans, Enterobacter cloacae, Bacillus sp., X. fastidiosa, Xanthomonas axonopodis pv. citri, and Candidatus Liberibacter asiaticus. We have demonstrated that with these methods we can quantitatively monitor the colonization of xylem by M. mesophilicum during the course of disease development in plants artificially inoculated with both bacteria.

Citrus sinensis↗
PubMed2004

Influence of resistance exercise training on glucose control in women with type 2 diabetes.

The objective of the study was to evaluate the effects of acute and chronic resistance training on glucose and insulin responses to a glucose load in women with type 2 diabetes. Subjects consisted of type 2 diabetic women (n = 7) and age-matched controls (n = 8) with normal glucose tolerance. All subjects participated in 3 oral glucose tolerance tests: pretraining, 12 to 24 hours after the first exercise session (acute) and 60 to 72 hours after the final training session (chronic). Exercise training consisted of a whole body resistance exercise program using weight-lifting machines 3 days per week for 6 weeks. Resistance training was effective in increasing strength of all muscle groups in all subjects. Integrated glucose concentration expressed as area under the curve (AUC) was 3,355.0 +/- 324.6 mmol/L. min pretraining, improved significantly (P <.01) after the acute bout of exercise (2,868 +/- 324.0 mmol/L. min), but was not improved with chronic training (3,206.0 +/- 337.0 mmol/L. min) in diabetic subjects. A similar pattern of significance was observed with peak glucose concentration (pre: 20.2 +/-1.4 mmol/L; acute: 17.2 +/- 1.7 mmol/L; chronic: 19.9 +/- 1.7 mmol/L). There were no significant changes in insulin concentrations after any exercise bout in the diabetic subjects. There were no changes in glucose or insulin levels in control subjects. An acute bout of resistance exercise was effective in improving integrated glucose concentration, including reducing peak glucose concentrations in women with type 2 diabetes, but not age-matched controls. There were no significant changes in insulin concentrations for either group. Resistance exercise offers an alternative to aerobic exercise for improving glucose control in diabetic patients. To realize optimal glucose control benefits, individuals must follow a regular schedule that includes daily exercise.

Adult↗
PubMed2004

Interaction between endophytic bacteria from citrus plants and the phytopathogenic bacteria Xylella fastidiosa, causal agent of citrus-variegated chlorosis.

AIMS: To isolate endophytic bacteria and Xylella fastidiosa and also to evaluate whether the bacterial endophyte community contributes to citrus-variegated chlorosis (CVC) status in sweet orange (Citrus sinensis [L.] Osbeck cv. Pera). METHODS AND RESULTS: The presence of Xylella fastidiosa and the population diversity of culturable endophytic bacteria in the leaves and branches of healthy, CVC-asymptomatic and CVC-symptomatic sweet orange plants and in tangerine (Citrus reticulata cv. Blanco) plants were assessed, and the in vitro interaction between endophytic bacteria and X. fastidiosa was investigated. There were significant differences in endophyte incidence between leaves and branches, and among healthy, CVC-asymptomatic and CVC-symptomatic plants. Bacteria identified as belonging to the genus Methylobacterium were isolated only from branches, mainly from those sampled from healthy and diseased plants, from which were also isolated X. fastidiosa. CONCLUSIONS: The in vitro interaction experiments indicated that the growth of X. fastidiosa was stimulated by endophytic Methylobacterium extorquens and inhibited by endophytic Curtobacterium flaccumfaciens. SIGNIFICANCE AND IMPACT OF THE STUDY: This work provides the first evidence of an interaction between citrus endophytic bacteria and X. fastidiosa and suggests a promising approach that can be used to better understand CVC disease.

Actinomycetales↗
PubMed2001

Resting leptin responses to acute and chronic resistance training in type 2 diabetic men and women.

PURPOSE: To evaluate the plasma leptin levels approximately 24 h post-exercise in control and type 2 diabetic subjects and to establish if observed changes in leptin concentrations were acute or chronic effects of a resistance training program. METHODS: Thirty men and women (17 controls and 13 type 2, obese diabetics, age 40-55 y) had resting blood samples drawn at 08:00 h (12 h postprandial) at the beginning of the study (pre-training), 24 h after a three repetition maximal weight lifting bout (acute) and 72 h after their last training bout of 6 weeks of resistance training (chronic). The two groups were not matched with respect to body mass index and the control subjects were not normal weight. Subjects weight-trained three times a week, for 6 weeks, for 1 h, training both the upper and lower body. RESULTS: Serum leptin concentrations were significantly higher in the type 2 diabetics than in the control group at pre-training (41.4+/-8.9 vs 11.4+/-3.0 ng/ml, P<0.05, respectively). Compared to pre-training, the leptin levels decreased significantly (P<0.01) after acute exercise in the diabetics but not in the control subjects (diabetics 30.9+/-7.1 vs controls 10.6+/-2.6 ng/ml). Approximately 72 h after 6 weeks of exercise training, the leptin concentrations were no longer lower than the pre-training values in either group (36.9+/-8.8 vs 11.9+/-8.8 ng/ml, respectively, P=NS). When leptin concentrations were log transformed and adjusted for fat mass there were still significant changes in leptin levels over time and between the control and diabetic group (P<0.05). CONCLUSIONS: The type 2 diabetics showed a significant 30% reduction in resting leptin levels 24 h after a single bout of resistance exercise. This was an acute response to resistance exercise and not a chronic training effect (no difference between pre-training and chronic). The decreased resting leptin concentrations approximately 24 h post-acute exercise may be due to reduced glucose availability to the adipose tissue, particularly in the diabetic subjects. There is no chronic effect of resistance exercise on leptin concentrations.

Adipose Tissue↗
PubMed2001

RAPD profile and antibiotic susceptibility of Xylella fastidiosa, causal agent of citrus variegated chlorosis.

AIMS: The aim of this study was to evaluate the diversity of Xylella fastidiosa isolated from citrus trees affected by Citrus Variegated Chlorosis (CVC). METHODS AND RESULTS: The antibiotic susceptibility by agar disc diffusion and minimum inhibitory concentration (MIC) methods was observed for all drug evaluated, except for penicillin-G. Genetic diversity by RAPD analysis revealed three major groups (citrus, coffee and grapevine), being the citrus group more similar with the coffee group than with the grapevine group. CONCLUSIONS, SIGNIFICANCE AND IMPACT OF THE STUDY: This study highlights the possibility to use these antibiotics susceptibility as markers in the development of a cloning vector and penicillin-G could be used as a selective marker for the isolation of X. fastidiosa from citrus plants.

Anti-Bacterial Agents↗
PubMed2001

Variability and interactions between endophytic bacteria and fungi isolated from leaf tissues of citrus rootstocks.

Fungi and bacteria were isolated from surface disinfected leaf tissues of several citrus rootstocks. The principal bacterial species isolated were Alcaligenes sp., Bacillus spp. (including B. cereus, B. lentus, B. megaterium, B. pumilus, and B. subtilis), Burkholderia cepacia, Curtobacterium flaccumfaciens, Enterobacter cloacae, Methylobacterium extorquens, and Pantoea agglomerans, with P. agglomerans and B. pumilus being the most frequently isolated species. The most abundant fungal species were Colletotrichum gloeosporioides, Guignardia citricarpa, and Cladosporium sp. Genetic variability between 36 endophytic bacterial isolates was analysed by the random amplified polymorphic DNA (RAPD) technique, which indicated that B. pumilus isolates were more diverse than P. agglomerans isolates, although genetic diversity was not related to the host plants. In vitro interaction studies between G. citricarpa isolates and the most frequently isolated endophytic bacteria showed that metabolites secreted by G. citricarpa have an inhibitory growth effect on some Bacillus species, and a stimulatory growth effect on P. agglomerans.

Ascomycota↗
PubMed2001

A biolistic process for in vitro gene transfer into chicken embryos.

Chicken embryos kept in culture medium were bombarded using a high helium gas pressure biolistic device. To optimize the factors that affect transformation efficiency, the lacZ gene under control of the human cytomegalovirus immediate early enhancer/promoter was used as a reporter gene. There was an inverse relationship between survival rate and transformation efficiency. The best conditions obtained for high embryo survival and high transformation efficiency were achieved with 800 psi helium gas pressure, 500 mmHg vacuum, gold particles, an 8 cm DNA-coated microparticle flying distance to the embryo and embryo placement 0.5 cm from the center of the particle dispersion cone. Under these conditions, transformation efficiency was 100%, survival rate 25% and the number of expression units in the embryo body cells ranged from 100 to 1,000. Expression of green fluorescent protein was also detected in embryos bombarded under optimal conditions. Based on the results obtained, the biolistic process can be considered an efficient method for the transformation of chicken embryos and therefore can be used as a model system to study transient gene expression and tissue-specific promoters.

Animals↗
PubMed2001

Pathogenicity of four strains of entomopathogenic fungi against the bovine tick Boophilus microplus.

OBJECTIVE: To evaluate the pathogenicity of 4 strains of the entomopathogenic fungi Metarhizium flavoviride var flavoviride and Metarhizium anisopliae var anisopliae in vitro against the bovine tick Boophilus microplus. SAMPLE POPULATION: 300 B. microplus females were used for each series of 5 conidial suspensions of the 4 fungal strains; a control group of 60 females was treated with sterilized water. PROCEDURE: For each fungal strain, sporulating cultures were used to produce conidial suspensions containing 10(4), 10(5), 10(6), 10(7), and 10(8) conidia/ml. A bioassay was performed by immersing B. microplus females in conidial suspensions; the procedure was repeated with a new group of 60 females for each dilution (a total of 300). A control group was immersed in the same volume of sterilized distilled water. Ticks were maintained at 27 C and 80% relative humidity for 7 days, and the following variables were evaluated: weight before egging, egging weight, percentage eclosion, reproductive efficiency, and percentage control. RESULTS: A low index of eclosion was found in the treated groups, with a progressive decrease in percentage eclosion (control = 100%) with increased conidial concentration. The highest level of effectiveness of biocontrol was detected in groups treated with concentrations of 10(7) and 10(8) conidia/ml. CONCLUSIONS AND CLINICAL RELEVANCE: Strains of the entomopathogenic fungi M. anisopliae var anisopliae and M. flavoviride var flavoviride were effective as biological control agents against B. microplus, with M. flavoviride strain CG-291 being the most effective. The pathogenicity of these strains and the development of an application method for this fungus for control of ticks may result in improved animal productivity and a reduction in environmental pollution through the use of fewer chemical insecticides.

Animals↗
PubMed2000

Protein kinase C modulates insulin action in human skeletal muscle.

There is good evidence from cell lines and rodents that elevated protein kinase C (PKC) overexpression/activity causes insulin resistance. Therefore, the present study determined the effects of PKC activation/inhibition on insulin-mediated glucose transport in incubated human skeletal muscle and primary adipocytes to discern a potential role for PKC in insulin action. Rectus abdominus muscle strips or adipocytes from obese, insulin-resistant, and insulin-sensitive patients were incubated in vitro under basal and insulin (100 nM)-stimulated conditions in the presence of GF 109203X (GF), a PKC inhibitor, or 12-deoxyphorbol 13-phenylacetate 20-acetate (dPPA), a PKC activator. PKC inhibition had no effect on basal glucose transport. GF increased (P < 0.05) insulin-stimulated 2-deoxyglucose (2-DOG) transport approximately twofold above basal. GF plus insulin also increased (P < 0.05) insulin receptor tyrosine phosphorylation 48% and phosphatidylinositol 3-kinase (PI 3-kinase) activity approximately 50% (P < 0.05) vs. insulin treatment alone. Similar results for GF on glucose uptake were observed in human primary adipocytes. Further support for the hypothesis that elevated PKC activity is related to insulin resistance comes from the finding that PKC activation by dPPA was associated with a 40% decrease (P < 0.05) in insulin-stimulated 2-DOG transport. Incubation of insulin-sensitive muscles with GF also resulted in enhanced insulin action ( approximately 3-fold above basal). These data demonstrate that certain PKC inhibitors augment insulin-mediated glucose uptake and suggest that PKC may modulate insulin action in human skeletal muscle.

Abdominal Muscles↗
PubMed1999

Dissemination of latent colon adenocarcinoma after laparoscopic cholecystectomy.

The seeding of neoplasms at trocar sites has been reported with increasing frequency. A case is presented of a 68 year-old woman with metastatic seeding of nondiagnosed colon cancer at the umbilical trocar site used for a laparoscopic cholecystectomy. Histopathological examination showed metastatic adenocarcinoma probably of gastrointestinal origin. A colonoscopy performed at the same time revealed a 2-cm lesion at the hepatic flexure which was shown histologically to be a differentiated adenocarcinoma. Diffuse peritoneal carcinomatosis was also found on computerised tomography. It is presumed that the primary colon cancer existed prior to cholecystectomy.

Adenocarcinoma↗
PubMed1998

Characterization of an Aspergillus nidulans mutant with abnormal distribution of nuclei in hyphae, metulae, phialides and conidia.

The V10 deteriorated variant of Aspergillus nidulans has hyphae, metulae, phialides and conidia with abnormal nuclear distributions. The alterations observed were: increase in the number of nuclei in hyphae, metulae and phialides, presence of anucleate, uninucleate and multinucleate conidia, abnormal vegetative growth and defection conidiation. When 0.5 M NaCl was added to the medium, an increase in the number of conidia was observed but their morphology and number of nuclei were not modified. The gene responsible for these alterations was named anuA1. The anuA1 gene is located on linkage group VII and is possibly involved in nuclear migration to hyphae, metulae, phialides and conidia.

Aspergillus nidulans↗
PubMed1998

Training decreases muscle glycogen turnover during exercise.

The present study was undertaken to determine the effects of endurance training on glycogen kinetics during exercise. A new model describing glycogen kinetics was applied to quantitate the rates of synthesis and degradation of glycogen. Trained and untrained rats were infused with a 25% glucose solution with 6-3H-glucose and U-14C-lactate at 1.5 and 0.5 microCi x min(-1) (where 1 Ci=3.7 x 10(10) Bq), respectively, during rest (30 min) and exercise (60 min). Blood samples were taken at 10-min intervals starting just prior to isotopic infusion, until the cessation of exercise. Tissues harvested after the cessation of exercise were muscle (soleus, deep, and superficial vastus lateralis, gastrocnemius), liver, and heart. Tissue glycogen was quantitated and analyzed for incorporation of 3H and 14C via liquid scintillation counting. There were no net decreases in muscle glycogen concentration from trained rats, whereas muscle glycogen concentration decreased to as much as 64% (P < 0.05) in soleus in muscles from untrained rats after exercise. Liver glycogen decreased in both trained (30%) and untrained (40%) rats. Glycogen specific activity increased in all tissues after exercise indicating isotope incorporation and, thus, glycogen synthesis during exercise. There were no differences in muscle glycogen synthesis rates between trained and untrained rats after exercise. However, training decreased muscle glycogen degradation rates in total muscle (i.e., the sum of the degradation rates of all of the muscles sampled) tenfold (P < 0.05). We have applied a model to describe glycogen kinetics in relation to glucose and lactate metabolism during exercise in trained and untrained rats. Training significantly decreases muscle glycogen degradation rates during exercise.

Animals↗
PubMed1997

Vanadate stimulation of 2-deoxyglucose transport is not mediated by PI 3-kinase in human skeletal muscle.

Glucose transport in mammalian skeletal muscle is stimulated by insulin, hypoxia and tyrosine protein phosphatase inhibitors such as vanadate. However, it is unknown whether the vanadate signaling mechanism shares a common or separate pathway with insulin or hypoxia. Therefore, experiments were conducted on incubated human muscle strips to compare the effects of vanadate with insulin and hypoxia stimulated 2-deoxyglucose transport (2-DOG). We also used the phosphatidylinositol 3-kinase (PI 3-kinase) inhibitor wortmannin to examine whether PI 3-kinase is a common step by which each stimulate glucose transport. Results demonstrate that whereas the effects of vanadate and hypoxia were additive with insulin stimulated glucose transport, the effect of vanadate plus hypoxia was not. In addition, wortmannin significantly (P < 0.05) reduced insulin but not vanadate or hypoxia stimulated 2-DOG transport. Moreover, PI 3-kinase activity was significantly elevated (P < 0.05) in the presence of insulin but not vanadate. In conclusion, these data suggest that vanadate and hypoxia stimulate glucose transport via a similar signaling pathway which is distinct from insulin and that the vanadate signaling pathway is not mediated by PI 3-kinase in human skeletal muscle.

Analysis of Variance↗
PubMed1997

Alterations in skeletal muscle protein-tyrosine phosphatase activity and expression in insulin-resistant human obesity and diabetes.

Obese human subjects have increased protein-tyrosine phosphatase (PTPase) activity in adipose tissue that can dephosphorylate and inactivate the insulin receptor kinase. To extend these findings to skeletal muscle, we measured PTPase activity in the skeletal muscle particulate fraction and cytosol from a series of lean controls, insulin-resistant obese (body mass index > 30) nondiabetic subjects, and obese individuals with non-insulin-dependent diabetes. PTPase activities in subcellular fractions from the nondiabetic obese subjects were increased to 140-170% of the level in lean controls (P < 0.05). In contrast, PTPase activity in both fractions from the obese subjects with non-insulin-dependent diabetes was significantly decreased to 39% of the level in controls (P < 0.05). By immunoblot analysis, leukocyte antigen related (LAR) and protein-tyrosine phosphatase 1B had the greatest increase (threefold) in the particulate fraction from obese, nondiabetic subjects, and immunodepletion of this fraction using an affinity-purified antibody directed at the cytoplasmic domain of leukocyte antigen related normalized the PTPase activity when compared to the activity from control subjects. These findings provide further support for negative regulation of insulin action by specific PTPases in the pathogenesis of insulin resistance in human obesity, while other regulatory mechanisms may be operative in the diabetic state.

Adult↗
PubMed1996

Similar evolutionary patterning among repeated and single copy nematode mitochondrial genes.

Ten of 12 mitochondrial protein-coding genes and the large (16S) mitochondrial rRNA gene have been identified and mapped within the Romanomermis culicivorax mitochondrial genome. This transcriptional map differs from other nematode mitochondrial DNAs (mtDNAs) with respect to gene order and transcriptional orientation of some genes. Several of these coding regions are components of a 3.0-kilobase mtDNA repeating unit, allowing a direct comparison of nucleotide and amino acid sequence composition for repeated and single copy genes. Analysis of protein-coding regions representing repeated (ND3, ND6) and single copy genes (ATPase 6, cyt.b, COI, COIII, ND1, ND4, ND5), and four repeat-associated open reading frames (ORFs) with unassigned function have revealed striking similarities in nucleotide composition, amino acid frequencies, and codon biases. Although we anticipated that reiterated protein coding regions might be evolving under relaxed selection, our results indicate that both repeated and unique mitochondrial genes appear subject to similar functional constraints.

Amino Acid Sequence↗
PubMed1995

Skeletal muscle fiber composition is related to adiposity and in vitro glucose transport rate in humans.

The purpose of this study was to determine if a relationship exists among skeletal muscle fiber composition, adiposity, and in vitro muscle glucose transport rate in humans. Rectus abdominus muscle was obtained during elective abdominal surgery from nonobese control (n = 12), obese (n = 12), and obese non-insulin-dependent diabetes mellitus (NIDDM) patients (n = 10). The obese NIDDM group had a significantly lower percentage of type I muscle fibers (32.2 +/- 1.9%) than the obese group (40.4 +/- 2.7%), and both obese groups were significantly lower than the control group (50.0 +/- 2.6%). Insulin-stimulated glucose transport, determined on 28 subjects, was significantly lower in both the obese (3.83 +/- 0.48 nmol.min-1.mg-1) and NIDDM (3.93 +/- 1.0 nmol.min-1.mg-1) groups vs. the control group (7.35 +/- 1.50 nmol.min-1.mg-1). Body mass index (BMI) was inversely correlated to percent type I fibers (r = -0.50, P < 0.01) and to the insulin-stimulated glucose transport rate (r = -0.53, P < 0.01). The percentage of type I muscle fibers was related to the insulin-stimulated glucose transport rate (r = 0.57, P < 0.01), although this relationship was not significant after adjusting for BMI. Although these data do not support an independent relationship between fiber type and insulin action in obesity, a reduced skeletal muscle type I fiber population may be one component of a multifactorial process involved in the development of insulin resistance.

Adipose Tissue↗