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Biomedical subjects

M A Machado

Publications and source records attributed to M A Machado.

At least 19 recordsLinked to original sources
PubMed2006

DGAT1 K232A polymorphism in Brazilian cattle breeds.

Recent reports identified DGAT1 (EC 2.3.1.20) harboring a lysine to alanine substitution (K232A) as a candidate gene with a strong effect on milk production traits. Our objective was to estimate the frequency of the DGAT1 K232A polymorphism in the main Zebu and Taurine breeds in Brazil as well as in Zebu x Taurine crossbreds as a potential QTL for marker-assisted selection. Samples of 331 animals from the main Brazilian breeds, Nellore, Guzerat, Red Sindhi, Gyr, Holstein, and Gyr x Holstein F1 were genotyped for DGAT1 K232A polymorphism (A and K alleles) using the PCR-RFLP technique. The highest frequency of the A allele was found in the Holstein sample (73%) followed by Gyr x Holstein F1 (39%). Gyr and Red Sindhi showed low frequencies of A alleles (4 and 2.5%, respectively). The A allele was not found in the Nellore and Guzerat samples. Our results could be used to guide association studies between this locus and milk traits in these breeds.

Animals↗
PubMed2006

Association of BoLA-DRB3.2 alleles with tick (Boophilus microplus) resistance in cattle.

Losses caused by bovine tick burdens in tropical countries have a tremendous economic impact on production systems. Besides reducing production, this parasite can cause death in the most susceptible animals. The use of commercial acaricides has been the major method of control, but their misuse has led to tick resistance to many chemicals. More recently, vaccines have been used in some countries without solving the problem completely. An alternative could be the development of resistant animals and the use of genetic markers and candidate genes that could help with the enormous task of selecting resistant animals. The bovine lymphocyte antigen genes (BoLA) have been shown to be associated with some parasitic infestations and disease incidence. Thus, the objective of the present study was to determine the association of BoLA-DRB3.2 alleles with tick resistance in cattle. The study was conducted on 231 F2 (Gyr x Holstein) animals that were artificially infested with 10,000 tick larvae. Log of tick count +1 was used as the dependent variable in a mixed animal model with allele substitution effects in addition to fixed effects of year and season at tick count, sex of calves, age of animal at tick count, hair type (short-straight, short-curl, long-straight, and long-curl), coat color (white, >75% white, 50- 75% white, and 25-50% white), and additive genetic, permanent environmental and residual effects as random. Females showed fewer ticks than males. Animals with short-straight hair were more resistant to tick infestation than animals with long-curl hair, and animals with whiter coat color also had fewer ticks. An association between BoLA alleles and lower tick number was found for alleles DRB3.2 *18, *20 and *27 at the 5% significance level. Also, one allele (DRB3.2*16) showed an association at the 10% level. Allele *27 was the most frequent in the population (30.7%), followed by alleles *16 (10.8%), *20 (8.7%) and *18 (2.4%). These results suggest that BoLA-DRB3.2 alleles could be used to help in the selection of animals resistant to tick infestation. However, further studies involving a larger population of cattle in combination with other BoLA genes may help to understand the mechanisms of resistance to parasites.

Alleles↗
PubMed2006

Complete nucleotide sequence, genomic organization and phylogenetic analysis of Citrus leprosis virus cytoplasmic type.

The complete nucleotide sequence of the genomic RNA 1 (8745 nt) and RNA 2 (4986 nt) of Citrus leprosis virus cytoplasmic type (CiLV-C) was determined using cloned cDNA. RNA 1 contains two open reading frames (ORFs), which correspond to 286 and 29 kDa proteins. The 286 kDa protein is a polyprotein putatively involved in virus replication, which contains four conserved domains: methyltransferase, protease, helicase and polymerase. RNA 2 contains four ORFs corresponding to 15, 61, 32 and 24 kDa proteins, respectively. The 32 kDa protein is apparently involved in cell-to-cell movement of the virus, but none of the other putative proteins exhibit any conserved domain. The 5' regions of the two genomic RNAs contain a 'cap' structure and poly(A) tails were identified in the 3'-terminals. Sequence analyses and searches for structural and non-structural protein similarities revealed conserved domains with members of the genera Furovirus, Bromovirus, Tobravirus and Tobamovirus, although phylogenetic analyses strongly suggest that CiLV-C is a member of a distinct, novel virus genus and family, and definitely demonstrate that it does not belong to the family Rhabdoviridae, as previously proposed. Based on these results it was proposed that Citrus leprosis virus be considered as the type member of a new genus of viruses, Cilevirus.

Base Sequence↗
PubMed2006

Primers based on the rpf gene region provide improved detection of Xanthomonas axonopodis pv. citri in naturally and artificially infected citrus plants.

AIMS: To have a PCR-based detection method for Xanthomonas axonopodis pv. citri (Xac) using primers designed in a specific region of its genome. METHODS AND RESULTS: A Xac-specific region was identified inside the rpf gene cluster of strain IAPAR 306 in an analysis of its complete genomic sequence. Two primers were designed, Xac01 and Xac02, which, when used in a standard PCR assay, direct the amplification of a 581 bp fragment from DNA of strains belonging to Xac from different regions around the world including unusual American and Asian strains. This product was not observed when DNA from strains of the closely related X. a. aurantifolli and X. a. citrumelo were used as templates. Extracts prepared from 28 xanthomonads of other species, and epiphytic bacteria isolated from citrus also failed to produce products with these primers. Amplification was obtained from cells grown in vitro, from extracts of both fresh and dried citrus canker lesions and from washes of inoculated but asymptomatic leaf surfaces. In sensitivity tests, this PCR technique detected as few as 100 cells. CONCLUSIONS: Primers Xac01 and Xac02 provide specific and sensitive detection of Xac in all citrus tissues where the pathogen is found. SIGNIFICANCE AND IMPACT OF THE STUDY: This PCR-based diagnostic test is suitable for monitoring asymptomatic plants in areas where the bacteria is endemic, in plant quarantine and regulatory situations, and also for obtaining an accurate diagnosis in a very short time. These are important characteristics for any assay to be used for the management of citrus canker disease.

Bacterial Infections↗
PubMed2004

Light/dark cycle-dependent metabolic changes in adipose tissue of pinealectomized rats.

We investigated the effects of pinealectomy on adipose tissue metabolism at different times of day. Adult male Wistar rats were divided into two groups: pinealectomized and control (sham-operated). Eight weeks after surgery, the animals were killed at three different times (at 8.00 a.m., at 4.00 p.m. and 11.00 p.m.). We collected blood samples for glucose, insulin, corticosterone, and leptin determinations, and periepididymal adipocytes for in vitro insulin-stimulated glucose uptake, oxidation, and incorporation into lipids. Pinealectomy caused insulin resistance as measured by 2-deoxyglucose uptake (a fall of approximately 40 % in the maximally insulin-stimulated rates) accompanied by hypercorticosteronemia at the three time points investigated without changes in plasma insulin an or leptin levels. Furthermore, pinealectomy increased the insulin-induced glucose incorporation into lipids (77 %) at 4.00 p.m. and insulin-induced glucose oxidation in the morning and in the afternoon, while higher rates were observed in the evening and in the morning in control rats. In conclusion, cell responsiveness to insulin was differentially affected by pineal ablation and time of day, and persistent insulin resistance was obtained in pinealectomized rats. We hypothesize that pinealectomy exposes the animal to an inadequate match between energy requirements and fuel mobilization.

Adipocytes↗
PubMed2003

Comparative analyses of the complete genome sequences of Pierce's disease and citrus variegated chlorosis strains of Xylella fastidiosa.

Xylella fastidiosa is a xylem-dwelling, insect-transmitted, gamma-proteobacterium that causes diseases in many plants, including grapevine, citrus, periwinkle, almond, oleander, and coffee. X. fastidiosa has an unusually broad host range, has an extensive geographical distribution throughout the American continent, and induces diverse disease phenotypes. Previous molecular analyses indicated three distinct groups of X. fastidiosa isolates that were expected to be genetically divergent. Here we report the genome sequence of X. fastidiosa (Temecula strain), isolated from a naturally infected grapevine with Pierce's disease (PD) in a wine-grape-growing region of California. Comparative analyses with a previously sequenced X. fastidiosa strain responsible for citrus variegated chlorosis (CVC) revealed that 98% of the PD X. fastidiosa Temecula genes are shared with the CVC X. fastidiosa strain 9a5c genes. Furthermore, the average amino acid identity of the open reading frames in the strains is 95.7%. Genomic differences are limited to phage-associated chromosomal rearrangements and deletions that also account for the strain-specific genes present in each genome. Genomic islands, one in each genome, were identified, and their presence in other X. fastidiosa strains was analyzed. We conclude that these two organisms have identical metabolic functions and are likely to use a common set of genes in plant colonization and pathogenesis, permitting convergence of functional genomic strategies.

Base Sequence↗
PubMed2002

Comparison of the genomes of two Xanthomonas pathogens with differing host specificities.

The genus Xanthomonas is a diverse and economically important group of bacterial phytopathogens, belonging to the gamma-subdivision of the Proteobacteria. Xanthomonas axonopodis pv. citri (Xac) causes citrus canker, which affects most commercial citrus cultivars, resulting in significant losses worldwide. Symptoms include canker lesions, leading to abscission of fruit and leaves and general tree decline. Xanthomonas campestris pv. campestris (Xcc) causes black rot, which affects crucifers such as Brassica and Arabidopsis. Symptoms include marginal leaf chlorosis and darkening of vascular tissue, accompanied by extensive wilting and necrosis. Xanthomonas campestris pv. campestris is grown commercially to produce the exopolysaccharide xanthan gum, which is used as a viscosifying and stabilizing agent in many industries. Here we report and compare the complete genome sequences of Xac and Xcc. Their distinct disease phenotypes and host ranges belie a high degree of similarity at the genomic level. More than 80% of genes are shared, and gene order is conserved along most of their respective chromosomes. We identified several groups of strain-specific genes, and on the basis of these groups we propose mechanisms that may explain the differing host specificities and pathogenic processes.

Gene Order↗
PubMed2001

Absorbable versus nonabsorbable membranes and bone grafts in the treatment of ligature-induced peri-implantitis defects in dogs. Part I. A clinical investigation.

The purpose of this study was to clinically evaluate an absorbable collagen membrane (Bio-Gide) and a nonabsorbable polytetrafluoroethylene membrane (PTFE), associated or not with bone grafts, for the treatment of ligature-induced peri-implantitis defects in dogs. The bilateral mandibular premolars were removed from 5 2-year-old mongrel dogs. After 3 months of healing, 3 titanium implants were placed on each side of the mandible. Experimental peri-implantitis was induced after abutment connection. Ligatures and abutments were removed after 1 month and the bone defects were randomly assigned to one of the following treatments: DB: debridement alone; GBR+BG-I: debridement plus PTFE membrane associated with mineralized bone graft (Bio-Oss); GBR+BG-II: debridement plus collagen membrane (Bio-Gide) associated with mineralized bone graft; GBR-I: debridement plus PTFE membrane; GBR-II: debridement plus collagen membrane; BG: debridement plus mineralized bone graft. The peri-implant bone defects were measured before and 5 months after treatment. Results showed the greatest percentage of vertical bone fill for GBR+BG-II (27.77+/-14.07) followed by GBR-II (21.78+/-16.19), BG (21.26+/-6.87), GBR+BG-I (19.57+/-13.36), GBR-I (18.86+/-10.63) and DB (14.03+/-5.6). However, the values were not statistically significant (ANOVA, contrast F test, P=0.612). Within the limits of the present investigation, it can be concluded that no difference was detected among treatments.

Absorbable Implants↗
PubMed2001

Clinical and microbiological evaluation of ligature-induced peri-implantitis and periodontitis in dogs.

The purpose of this study was to evaluate the attachment loss around teeth and implants by clinical and microbiological analysis. The mandibular premolars were extracted in 5 mongrel dogs and, 3 months later, two titanium implants were installed on each side of the mandible and, after another 3 months, abutment connection was performed. Plaque control in the implants and maxillary premolars was maintained for two weeks prior to the start of the main experiment. On day 0 and 30 days after ligature placement, microbiological samples were obtained and relative attachment level was measured for the teeth and implants. The presence of Porphyromonas gingivalis, Bacteroides forsythus, Actinobacillus actinomycetemcomitans, Prevotella intermedia and Prevotella nigrescens was evaluated by polymerase chain reaction technique on day 0 and 30 days after ligature placement. None of the above bacteria were detected on day 0. Thirty days after ligature placement, P. gingivalis was present in 95% and 85% and B. forsythus was present in 80% and 85% of the implants and teeth sites, respectively. Statistical analysis (one-way RM-ANOVA) showed a significant difference (P<0.01) between pre- and post-induction measurements around teeth and implants. However, there was no significant difference (P=0.41) in the rate of attachment loss, between periodontitis and peri-implantitis. It can be concluded that: (1) P. gingivalis and B. forsythus were strongly associated with induced peri-implantitis and periodontitis, and (2) induced peri-implantitis and periodontitis presented a similar rate of attachment loss.

Animals↗
PubMed2001

Histometric evaluation of the effect of nicotine administration on periodontal breakdown: an in vivo study.

The present study investigated the effect of nicotine administration on periodontal breakdown resulting from ligature-induced periodontitis in rats. Twenty adult male Wistar rats were used. After anesthesia, a mandibular first molar was randomly assigned to receive a cotton ligature in the sulcular area while the contralateral tooth was left unligated. The animals were randomly assigned to one of the following treatments. of daily intraperitoneal injections: A - saline solution, B -0.37 mg of nicotine kg, C -0.57 mg of nicotine kg and D -0.73 mg of nicotine/kg. Thirty days later, the animals were sacrificed and the specimens routinely processed for serial decalcified sections. Statistical analysis (ANOVA) revealed greater bone loss (p<0.05) in the ligated teeth of animals which received nicotine (groups B/C D) than in the ligated teeth of animals which received saline solution (group A). In addition, a dose-dependent response was observed among the nicotine groups. A negative effect of nicotine was observed in the unligated teeth of the experimental groups (p<0.05). Therefore, daily administration of nicotine enhanced, in a dose-dependent manner, the effects of local factors in producing periodontal breakdown. Furthermore, the nicotine seemed to have a direct deleterious effect on the periodontal tissues.

Alveolar Bone Loss↗
PubMed2001

Differentiation of strains of Xylella fastidiosa by a variable number of tandem repeat analysis.

Short sequence repeats (SSRs) with a potential variable number of tandem repeat (VNTR) loci were identified in the genome of the citrus pathogen Xylella fastidiosa and used for typing studies. Although mono- and dinucleotide repeats were absent, we found several intermediate-length 7-, 8-, and 9-nucleotide repeats, which we examined for allelic polymorphisms using PCR. Five genuine VNTR loci were highly polymorphic within a set of 27 X. fastidiosa strains from different hosts. The highest average Nei's measure of genetic diversity (H) estimated for VNTR loci was 0.51, compared to 0.17 derived from randomly amplified polymorphic DNA (RAPD) analysis. For citrus X. fastidiosa strains, some specific VNTR loci had a H value of 0.83, while the maximum value given by specific RAPD loci was 0.12. Our approach using VNTR markers provides a high-resolution tool for epidemiological, genetic, and ecological analysis of citrus-specific X. fastidiosa strains.

Bacterial Typing Techniques↗
PubMed2001

[EPI-FLAIR sequence in the evaluation of mesial temporal sclerosis].

The purpose of this study is to evaluate morphological and signal intensity changes in the hippocampus in patients with medically intractable temporal lobe epilepsy. We studied 8 patients with mesial temporal sclerosis using a 1.5 -T MR and the following sequences Spin Eco- SE, Fast Spin Echo- FSE, Fluid Attenuation Inversion Recovery Echo Planar Imaging--FLAIR-EPI. We noticed a sensitive increase signal intensity on FLAIR- EPI sequences, particularly, in coronal images, than on SE and FSE sequences. The STIR sequence showed a cortical hippocampus atrophy in half of the cases, in whom signal abnormalities were present.

Echo-Planar Imaging↗
PubMed2001

Lack of evidence for superantigen activity of Toxoplasma gondii towards human T cells.

Toxoplasma gondii is an obligatory intracellular parasite whose life cycle may include man as an intermediate host. More than 500 million people are infected with this parasite worldwide. It has been previously reported that T. gondii contains a superantigen activity. The purpose of the present study was to determine if the putative superantigen activity of T. gondii would manifest towards human T cells. Peripheral blood mononuclear cells (PBMC) from individuals with no previous contact with the parasite were evaluated for proliferation as well as specific Vbeta expansion after exposure to Toxoplasma antigens. Likewise, PBMC from individuals with the congenital infection were evaluated for putative Vbeta family deletions in their T cell repertoire. We also evaluated, over a period of one year, the PBMC proliferation pattern in response to Toxoplasma antigens in patients with recently acquired infection. Some degree of proliferation in response to T. gondii was observed in the PBMC from individuals never exposed to the parasite, accompanied by specific Vbeta expansion, suggesting a superantigen effect. However, we found no specific deletion of Vbeta (or Valpha) families in the blood of congenitally infected individuals. Furthermore, PBMC from recently infected individuals followed up over a period of one year did not present a reduction of the Vbeta families that were originally expanded in response to the parasite antigens. Taken together, our data suggest that T. gondii does not have a strong superantigen activity on human T cells.

Adult↗
PubMed2001

[Dilatation of Virchow-Robin spaces in patients with migraine].

The Virchow-Robin spaces (VRS) are subpial invaginations making a tubular revestment scabbard that contains a vase between the subarachnoid and subpial spaces. The magnetic resonance imaging (MRI) is the only imaging method able for evaluate this anatomic detail. We studied the possible association between dilated VRS and migraine. Seventy patients with ages ranging from 13 to 54 years (mean, 36.5 years), with clinical diagnosis of migraine were studied and compared with a control group, without past of headache, composed by the same number of patients with ages ranged from 14 to 64 years (mean, 42 years). We observed widening of VRS in 28 cases (40%) of the migraine patients. In the control group this feature was demonstrated in only 5 cases (7.1%). Alert for the importance of detecting and recognizing the VRS, as well as, the differentiation from lacunar infarctions and choroid fissure cysts.

Adolescent↗
PubMed2001

Clinical study of guided bone regeneration and/or bone grafts in the treatment of ligature-induced peri-implantitis defects in dogs.

This study evaluated, by clinical analysis, the hard tissue response following treatment of ligature-induced peri-implantitis defects in 5 dogs. The mandibular premolars were removed from both sides of the jaw. After 3 months of healing, two titanium implants were placed on each side of the mandible. Following abutment connection, 3 months later, experimental peri-implantitis was induced by the placement of cotton ligatures in a submarginal position. Ligatures and abutments were removed after one month and the bony defects were randomly assigned to one of the following treatments: debridement (DE), debridement plus guided bone regeneration (GBR), debridement plus mineralized bone graft (BG) and debridement plus guided bone regeneration associated with mineralized bone graft (GBR + BG). The peri-implant bone defects were clinically measured before and 5 months post-treatment. Results showed a higher percentage of vertical bone fill for GBR + BG (27.77 +/- 14.07) followed by GBR (21.78 +/- 16.19), BG (21.26 +/- 6.87), DE (14.03 +/- 5.6). However, there were no statistically significant differences between any of the treatments proposed (one way repeated measures analysis of variance, P = 0.265).

Alveolar Bone Loss↗
PubMed2000

The genome sequence of the plant pathogen Xylella fastidiosa. The Xylella fastidiosa Consortium of the Organization for Nucleotide Sequencing and Analysis.

Xylella fastidiosa is a fastidious, xylem-limited bacterium that causes a range of economically important plant diseases. Here we report the complete genome sequence of X. fastidiosa clone 9a5c, which causes citrus variegated chlorosis--a serious disease of orange trees. The genome comprises a 52.7% GC-rich 2,679,305-base-pair (bp) circular chromosome and two plasmids of 51,158 bp and 1,285 bp. We can assign putative functions to 47% of the 2,904 predicted coding regions. Efficient metabolic functions are predicted, with sugars as the principal energy and carbon source, supporting existence in the nutrient-poor xylem sap. The mechanisms associated with pathogenicity and virulence involve toxins, antibiotics and ion sequestration systems, as well as bacterium-bacterium and bacterium-host interactions mediated by a range of proteins. Orthologues of some of these proteins have only been identified in animal and human pathogens; their presence in X. fastidiosa indicates that the molecular basis for bacterial pathogenicity is both conserved and independent of host. At least 83 genes are bacteriophage-derived and include virulence-associated genes from other bacteria, providing direct evidence of phage-mediated horizontal gene transfer.

Bacterial Adhesion↗