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Biomedical subjects

J Silva

Publications and source records attributed to J Silva.

At least 19 recordsLinked to original sources

Evaluation of the energy expenditure in competitive swimming strokes.

The purpose of this study was to measure and compare the total energy expenditure of the four competitive swimming strokes. Twenty-six swimmers of international level were submitted to an incremental set of 200-m swims (5 swimmers at Breaststroke, 5 swimmers at Backstroke, 4 swimmers at Butterfly and 12 swimmers at Front Crawl). The starting velocity was approximately 0.3 m x s (-1) less than a swimmer's best performance and thereafter increased by 0.05 m x s (-1) after each swim until exhaustion. Cardio-pulmonary and gas exchange parameters were measured breath-by-breath (BxB) for each swim to analyze oxygen consumption (VO2) and other energetic parameters by portable metabolic cart (K4b(2), Cosmed, Rome, Italy). A respiratory snorkel and valve system with low hydrodynamic resistance was used to measure pulmonary ventilation and to collect breathing air samples. Blood samples from the ear lobe were collected before and after each swim to analyze blood lactate concentration (YSI 1500 L, Yellow Springs, Ohio, USA). Total energy expenditure (E(tot)), was calculated for each 200-m stage. E (tot) differed significantly between the strokes at all selected velocities. At the velocity of 1.0 m x s (-1) and of 1.2 m x s (-1) the E(tot) was significantly higher in Breaststroke than in Backstroke, in Breaststroke than in Freestyle and in Butterfly than in Freestyle. At the velocity of 1.4 m x s (-1), the E(tot) was significantly higher in Breaststroke than in Backstroke, in Backstroke than in Freestyle, in Breaststroke than in Freestyle and in Butterfly than in Freestyle. At the velocity of 1.6 m x s (-1), the E(tot) was significantly higher in Breaststroke and in Butterfly than in Freestyle. As a conclusion, E(tot) of well-trained competitive swimmers was measured over a large range of velocities utilising a new BxB technique. Freestyle was shown to be the most economic among the competitive swimming strokes, followed by the Backstroke, the Butterfly and the Breaststroke.

Analysis of Variance↗

The presence of an intronic deletion in p73 and high levels of ZEB1 alter the TAp73/DeltaTAp73 ratio in colorectal carcinomas.

TAp73 variants largely mimic p53 suppressor activities, while DeltaTAp73 forms act as oncogenes through the inactivation of p53 and TAp73. The present study analysed how TAp73 and DeltaTAp73 levels might be affected by the presence of a 73 bp deletion in a regulatory region of p73. The clinical relevance of this deletion was also examined. ZEB1 can bind to the region repressing p73 transcription in vitro. The relationship between ZEB1 and p73 variant expression levels was studied in the context of this deletion and the levels of the ZEB1 cofactors p300 and CtBP. Tumour and normal tissue from 81 colorectal cancer patients was analysed to evaluate firstly the levels of TAp73, DeltaTAp73 (DeltaEx2p73, DeltaEx2/3p73, and DeltaNp73), ZEB1, p300, and CtBP by quantitative real-time RT-PCR, and secondly the presence of the 73 bp deletion. Tumour characteristics were examined in each patient. Suppressor and oncogenic isoforms of p73 were co-up-regulated in tumour tissues. Overexpression of p73 variants was associated with adverse tumour features. The 73 bp deletion was present in 40% of the patients and was associated with adverse pathological parameters of the tumours and also with TAp73 down-regulation. In those cases harbouring the deletion, the levels of ZEB1 and those of DeltaEx2p73, DeltaEx2/3p73, and DeltaNp73 correlated directly. Variations in the concentration of p300 affected the observed correlations between ZEB1 and the different p73 variants. In conclusion, in colorectal cancer, the 73 bp deletion in the first intron of the p73 gene and different expression levels of ZEB1 and p300 may act in concert to affect the ratio of TAp73/DeltaTAp73 forms, favouring p73 oncogenic variants. In addition, up-regulation of p73 oncogenic isoforms predicts a poor prognosis based on its relationship with advanced tumour stage.

Aged↗

A novel isoform of prostate apoptosis response 4 (PAR-4) that co-distributes with F-actin and prevents apoptosis in neural stem cells.

The elevated expression of prostate apoptosis response-4 (PAR-4) induces apoptosis in differentiating mouse embryonic stem (ES) cells. In embryoid body (EB) cells and the E15.5 stage of embryonic mouse brain, PAR-4 is expressed as two isoforms (38 and 33 kDa). Using mouse EB-derived RNA as a template we have cloned and characterized a novel isoform of PAR-4 (PAR-4/p33) that lacks exon 3 and shows a bona fide splice junction of exons 2 and 4. The molecular mass for PAR-4/p33 is estimated to be 33 kDa, corresponding to the short form found in the EB cells and E15.5 mouse brain. The fluorescent fusion protein of PAR-4/p33 is mainly found in the cytosol and is co-distributed with F-actin filaments, while that of the 38 kDa full length PAR-4/p38 is predominantly translocated to the nucleus. In contrast to the full length PAR-4 (PAR-4/p38), ectopic expression of PAR-4/p33 does not result in the activation of caspase 3 and the induction of apoptosis. PAR-4/p33 forms a complex with PAR-4/p38, which inhibits its nuclear translocation and the induction of apoptosis. PAR-4/p33 is suggested to be a dominant negative isoform of PAR-4/p38 and may regulate PAR-4-dependent apoptosis.

Actins↗

Characterization of the Bacteroides fragilis pathogenicity island in human blood culture isolates.

Bacteroides fragilis is an important anaerobic pathogen accounting for up to 10% of bacteremias in adult patients. Enterotoxin producing B. fragilis (ETBF) strains have been identified as enteric pathogens of children and adults. In order to further characterize the B. fragilis pathogenicity island (BfPAI) and using PCR assays for bft- and mpII-metalloprotease genes, we determined the frequency of B. fragilis strains with pattern I (containing the BfPAI and its flanking region), pattern II (lacking both the BfPAI and the flanking region), and pattern III (lacking the BfPAI but containing the flanking region) in 63 blood culture isolates. The results were compared to 197 B. fragilis isolates from different clinical sources. We found 19% of blood culture isolates were pattern I (ETBF), 43% were pattern II (NTBF) and 38% were pattern III (NTBF). Comparatively, B. fragilis isolates from other clinical sources were 10% pattern I, 47% pattern II and 43% pattern III. This suggests that the pathogenicity island and the flanking elements may be general virulence factors of B. fragilis.

Bacteremia↗

Hyperglycemia induced by pharmacological activation of central serotonergic pathways depends on the functional integrity of brain CRH system and 5-HT3 receptors.

In the present study, we investigated the effect of central serotonergic pathway activation achieved through third ventricle injections of quipazine, a serotonergic agonist, on plasma glucose levels of fasted and fed adult Wistar male rats, whose third ventricles were canulated 7 days before the experiments. Central quipazine administration induced a significant increase in plasma glucose levels in fasted animals, but was unable to modify plasma glucose concentrations in fed rats. Pretreatment with alpha-helical CRH, a CRH antagonist, significantly attenuated quipazine-induced hyperglycemia. Pretreatment with two different 5-HT3 receptor antagonists, LY-278,584 and ondansetron, was also able to produce a significant reduction in the hyperglycemic response evoked by central administration of quipazine. None of the antagonists used was capable of modifying plasma glucose concentrations when injected alone into the third ventricle. Quipazine-treated, hyperglycemic animals did not show any increase in plasma insulin levels. We conclude that acute pharmacological serotonergic stimulation by quipazine produces hyperglycemia by mechanisms that require the functional integrity of both CRH and 5-HT3 receptors, and that impairment in insulin secretion and/or activity may explain hyperglycemia induced by third ventricle injections of quipazine.

Animals↗

In situ gamma spectrometry measurements and Monte Carlo computations for the detection of radioactive sources in scrap metal.

A very limited number of field experiments have been performed to assess the relative radiation detection sensitivities of commercially available equipment used to detect radioactive sources in recycled metal scrap. Such experiments require the cooperation and commitment of considerable resources on the part of vendors of the radiation detection systems and the cooperation of a steel mill or scrap processing facility. The results will unavoidably be specific to the equipment tested at the time, the characteristics of the scrap metal involved in the tests, and to the specific configurations of the scrap containers. Given these limitations, the use of computer simulation for this purpose would be a desirable alternative. With this in mind, this study sought to determine whether Monte Carlo simulation of photon flux energy distributions resulting from a radiation source in metal scrap would be realistic. In the present work, experimental and simulated photon flux energy distributions in the outer part of a truck due to the presence of embedded radioactive sources in the scrap metal load are compared. The experimental photon fluxes are deduced by in situ gamma spectrometry measurements with portable Ge detector and the calculated ones by Monte Carlo simulations with the MCNP code. The good agreement between simulated and measured photon flux energy distributions indicate that the results obtained by the Monte Carlo simulations are realistic.

Computer Simulation↗

Effect of the pH of growth on the survival of Lactobacillus delbrueckii subsp. bulgaricus to stress conditions during spray-drying.

AIMS: The aim of this study was to optimize survival of Lactobacillus delbrueckii subsp. bulgaricus during spray-drying and subsequent storage through optimizing the pH of growth conditions. METHODS AND RESULTS: Cell concentrates previously grown without or with pH controlled were spray-dried and stored at 20 degrees C and heat treated at 57 degrees C. Cells grown under noncontrolled pH were more resistant to both drying and heating than cells grown under controlled pH but no significant differences were observed during storage. The intracellular proteins profile of cells grown under both conditions was studied by two-dimensional SDS-polyacrylamide gel electrophoresis. Eight proteins were identified using automated mass spectrometry (MS) and tandem mass spectrometry (MS/MS) data acquisition. Of the identified proteins, only cochaperonin GroES corresponded to a known heat shock protein (HSP). The other proteins identified are proteins involved in glycolysis. For cells grown under noncontrolled pH the expression of the Hsp70, GroES and GroEL, measured by Western blotting, was enhanced. CONCLUSIONS: The higher resistance of cells grown under noncontrolled pH correlates with the enhanced production of heat shock proteins. SIGNIFICANCE AND IMPACT OF THE STUDY: Growth of L. bulgaricus under controlled pH (commonly used by the starter cultures production industry) results in cells more sensitive to stresses frequently encountered by the cells during starter cultures preparation/storage/utilization.

Bacterial Proteins↗

Antimicrobial resistance and characterization of Helicobacter pylori strains isolated from Mexican adults with clinical outcome.

Eradication of Helicobacter pylori infection in Mexico is of great importance due to the elevated seroprevalence, however, there is yet very little information about antibiotic resistance rates in H. pylori isolates in our country. We analyzed susceptibility to three antimicrobials used in therapy of 49 H. pylori strains isolated from patients with active chronic gastritis, active chronic gastritis with lymphoid follicles, intestinal metaplasia and gastric cancer. All isolated strains were susceptible to amoxicillin, 28 (58%) were resistant to metronidazole and 2 (4%) were resistant to both clarithromycin and metronidazole. Sequence analysis of the 23S rRNA of the two clarithromycin-resistant strains showed the A2142G mutation in one and A2143G and T2182C mutations in the other. Metronidazole resistance was associated with cagA negative strains with a frequency of 82% (9/11). No significant correlation was found between vacA s/m alleles and metronidazole resistance.

Adult↗

Comparison of two major emergency department-based free-text chief-complaint coding systems.

INTRODUCTION: Emergency departments (EDs) using free-text chief-complaint data for syndromic surveillance face a unique challenge because a complaint might be described and coded in multiple ways. OBJECTIVE: Two major ED-based free-text chief-complaint coding systems were compared for agreement between free-text interpretation and syndrome coding. METHODS: Chief-complaint data from 21,736 patients at an urban ED were processed through both the New York City Department of Health and Mental Hygiene (DOHMH) syndrome coding system as modified by the Chicago Department of Public Health and the Real-Time Outbreak Detection System Complaint Coder (CoCo, version 2.1, University of Pittsburgh). To account for differences in each system's specified syndromes, relevant syndromes from the DOHMH system were collapsed into the corresponding CoCo categories so that a descriptive comparison could be made. DOHMH classifications were combined to match existing CoCo categories as follows: 1) vomit+diarrhea = Gastrointestinal; 2) cold+respiratory+asthma = Respiratory; 3) fevflu = Constitutional; 4) rash = Rash; 5) sepsis+other = Other, 6) unknown = Unknown. RESULTS: Overall agreement between DOHMH and CoCo syndrome coding was optimal (0.614 kappa). However, agreement between individual syndromes varied substantially. Rash and Respiratory had the highest agreement (0.711 and 0.594 kappa, respectively). Other and Constitutional had an intermediate level of agreement (0.453 and 0.419 kappa, respectively), but less than optimal agreement was identified for Gastrointestinal and Unknown (0.270 and 0.002 kappa, respectively). CONCLUSIONS: Although this analysis revealed optimal overall agreement between the two systems evaluated, substantial differences in classification schemes existed, highlighting the need for a consensus regarding chief-complaint classification.

Algorithms↗

SARS Surveillance Project--Internet-enabled multiregion surveillance for rapidly emerging disease.

INTRODUCTION: On March 15, 2003, CDC requested health-care and public health agencies to conduct surveillance for severe acute respiratory syndrome (SARS). The SARS Surveillance Project (SARS-SP) was established to rapidly implement multiregional SARS surveillance in emergency departments (EDs) by using existing Internet-based tools. OBJECTIVES: The objectives of SARS-SP were to 1) disseminate and update SARS screening forms for ED triage, 2) establish surveillance for SARS syndrome elements by using Regional Emergency Medicine Internet (REMI), 3) expand surveillance to multiple regions, and 4) evaluate the usefulness of Internet tools for agile surveillance during a rapidly emerging global epidemic. METHODS: SARS-SP developed, distributed, and updated an Internet-based triage form to identify patients for infection control and public health reporting. EDs then were invited to report visit frequencies with various SARS syndrome elements to local public health authorities by using the REMI Internet application (first in one metropolitan area, and later in four). After pilot-testing in one metropolitan area, the surveillance system was implemented in three others. RESULTS: Active syndromic surveillance was established by health departments in Milwaukee, Wisconsin; Denver, Colorado; Akron, Ohio; and Fort Worth, Texas. A total of 27 EDs reported syndrome frequencies from >146,000 patient encounters. CONCLUSIONS: ED and public health partners reported being satisfied with the system, confirming the usefulness of Internet tools in the rapid establishment of multiregion syndromic surveillance during an emerging global epidemic.

Communicable Diseases, Emerging↗

AZF and DAZ gene copy-specific deletion analysis in maturation arrest and Sertoli cell-only syndrome.

Deletions of the AZFc region in Yq11.2, which include the DAZ gene family, are responsible for most cases of male infertility and were associated with severe oligozoospermia and also with a variable testicular pathology. To uncover the functional contribution of DAZ to human spermatogenesis, a DAZ gene copy-specific deletion analysis was previously established and showed that DAZ1/DAZ2 deletions associate with oligozoospermia. In this study we applied the same screening method to 50 control fertile males and 91 non-obstructive azoospermic males, 39 with Sertoli cell-only syndrome (SCOS) and 52 with meiotic arrest (MA). Samples were also screened with 24 sequence-tagged sites to the different AZF regions, including 114 control fertile males. After biopsy (testicular sperm extraction, TESE), residual spermiogenesis was found in 57.7% MA and 30.8% SCOS cases (incomplete syndromes). DAZ1/DAZ2 deletions were associated with the testicular phenotype of residual spermiogenesis as they were only found in two patients (8%) with incomplete MA. Differences between incomplete (23.3%) and complete (4.5%) MA cases regarding AZFc and DAZ1/DAZ2 deletion frequencies, and between incomplete (58.3%) and complete (11.1%) SCOS cases for AZFc deletions, suggest that incomplete syndromes might represent an aggravation of the oligozoospermic phenotype. As successful TESE was achieved in 87.5% of MA cases with AZFc and DAZ1/DAZ2 deletions and in 58.3% of SCOS cases with AZFc deletions, the present results also suggest that these molecular markers might be used for the establishment of a prognosis before TESE.

Deleted in Azoospermia 1 Protein↗

Cloning and expression of Clostridium difficile toxin A gene (tcdA) by PCR amplification and use of an expression vector.

Toxigenic Clostridium difficile isolates harbor a 19 kb pathogenicity locus that encodes the genes for toxins A and B. Toxins A and B are among the largest known bacterial toxins expressing potent cytotoxicity and enterotoxicity, and thus the major virulence factors in C. difficile associated diarrhea. Cloning and sequencing of toxin genes is of interest for studies of molecular pathogenesis. We report the amplification and cloning of the complete toxin A gene into an Escherichia coli expression vector. Ten clones analyzed contained the complete toxin A gene. Four of these clones showed cytotoxic activity in cell culture, and were positive for toxin A as determined by ELISA. Toxin A expression was confirmed by Western immunoblot analysis. The presence of cytotoxic activity in cell culture suggests that toxin A activity is independent of other genes in the pathogenicity locus.

Bacterial Toxins↗

An abnormal screening ultrasound: concordance with a tertiary obstetrical ultrasound unit.

OBJECTIVE: To determine concordance of ultrasound diagnosis in referrals to a tertiary obstetrical ultrasound unit (TOU) for suspected abnormalities. STUDY DESIGN: Consecutive referrals for "abnormal outside ultrasound" during a 6-month period were compared with the TOU ultrasound diagnosis. Concordance of diagnosis was compared on the basis of organ system involved and referral for single or multiple suspected abnormalities. Chi(2) analysis was used; p < 0.05 was considered significant. RESULTS: Of 104 consecutive referrals reviewed, 42 (40.4%) had no abnormality documented at the TOU. Of the 62 abnormal ultrasound scans at the TOU, 78.3% were concordant. Concordance based on organ system involvement was central nervous system, 30.3%; cardiothoracic, 66.7%; gastrointestinal, 63.6%; genitourinary, 50%. Referrals for a single suspected anomaly were statistically no more likely to have a normal TOU ultrasound scan (40.4%) than those referred for multiple suspected anomalies (36.4%, p > 0.05). CONCLUSIONS: Most referrals to a tertiary center for "abnormal outside ultrasound" will be diagnosed with an abnormality.

Congenital Abnormalities↗

NF-kappa B inhibition markedly enhances sensitivity of resistant breast cancer tumor cells to tamoxifen.

Studies show that high Akt activity in breast carcinoma is associated with endocrine therapy resistance. Breast cancer cell lines expressing a constitutively active Akt are able to proliferate under reduced estrogen conditions, and are resistant to the growth inhibitory effects of tamoxifen. Understanding the targets of Akt signaling mediating tamoxifen resistance is of clinical significance. One possible target is nuclear factor kappa B (NF-kappa B), a transcription factor that plays a critical role in resistance to apoptosis and the induction of angiogenesis and invasion. In the present study, we found that Akt activity correlated with phosphorylation of I kappa B (the negative regulator of NF-kappa B), NF-kappa B DNA binding and tamoxifen resistance in vivo. Importantly, we found that co-treatment with the NF-kappa B inhibitor, parthenolide, or overexpression of I kappa B superrepressor restored tamoxifen sensitivity to our refractory Akt MCF-7 cells. These data suggest that activation of NF-kappa B via the PI3K/Akt signaling pathway may be a significant mechanism for development of endocrine therapy resistance in breast cancer, and that inhibition of NF-kappa B may be an effective treatment strategy to limit the progression of this disease.

Antineoplastic Agents, Hormonal↗

Experimental vitrification of human compacted morulae and early blastocysts using fine diameter plastic micropipettes.

BACKGROUND: Vitrification of human blastocysts has been successfully applied using grids, straws and cryoloops. We assessed the survival rate of human compacted morulae and early blastocysts vitrified in pipette tips with a smaller inner diameter and solution volume than the previously described open pulled straw (OPS) method. METHODS: Excess day 5 human embryos (n = 63) were experimentally vitrified in vessels. Embryos were incubated at 37 degrees C with sperm preparation medium (SPM) for 1 min, SPM + 7.5% ethylene glycol (EG)/dimethylsulphoxide (DMSO) for 3 min, and SPM + 16.5% EG + 16.5% DMSO + 0.67 mol/l sucrose for 25 s. They were then aspirated (0.5 microl) into a plastic micropipette tip (0.36 mm inner diameter), exposed to liquid nitrogen (LN(2)) vapour for 2 min before being placed into a pre-cooled cryotube, which was then closed and plunged into LN(2). Embryos were warmed and diluted using 0.33 mol/l and 0.2 mol/l sucrose. RESULTS: The survival rate for compacted morulae was 73% (22/30) and 82% (27/33) for early blastocysts. CONCLUSIONS: The survival rates of human compacted morulae and early blastocysts after vitrification with this simple technique are similar to those reported in the literature achieved by slow cooling and other vitrification protocols.

Blastocyst↗

Molecular characterization of beta-lactam resistance of Salmonella isolates from pediatric patients in Romania.

The molecular characterization of 16 clinical isolates of Salmonella enterica (14 serotype Typhimurium and 2 serotype Kingston) obtained between January and June 1999 from feces of children hospitalized in Iasi, Romania were genotypically compared by pulse field gel electrophoresis of XbaI restricted bacterial DNA. The majority of the clinical isolates (12/16) belonged to cluster A and (4/16) to unrelated strains, correlating to the OMP profile. Two major different patterns of beta-lactamases were identified: the first with pI of 5.4, 8.2 in 6/16 strains and the second with pI of 5.4 in 5/16. The blaTEM beta-lactamase was identified in 14/16 of the clinical isolates and the blaSHV-5 gene in one strain. We concluded that extended spectrum beta-lactamase (ESBL) with pIs of 8.2 was the most frequent enzyme produced by serotype Typhimurium isolates which were related.

Base Sequence↗

Systematic characterisation of silicon-embedded accelerometers for mechanomyography.

Silicon soft suction sockets (roll-on sleeves) currently used in passive prostheses for below-elbow amputees could also be used in externally powered prostheses, enhancing their functionality and comfort. However, as it is extremely difficult to hold currently used electromyography (EMG) sensors in place reliably within a silicon socket, an alternative measurement of muscular activity as the control input is necessary. Mechanomyography (MMG) is the epidermal measurement of the low-frequency vibrations produced by a contracting muscle. MMG sensors do not have to be in direct contact with the skin. Moreover, the embedding of sensors in the roll-on sleeve may also solve attachment issues, making sensor placement flexible. Therefore the objective was to determine the feasibility of recording MMG signals using silicon-embedded, micro-machined accelerometers. Fifteen embedded accelerometers were excited with predefined vibration patterns. The signal-to-noise ratio (SNR) and frequency response of each sample were measured and compared with those of non-embedded accelerometers. The SNR of embedded samples (approximately equal to 19 dB) was significantly higher than that of non-embedded samples (approximately equal to 12 dB), owing to the considerable mechanical damping effect of the silicon in the 300-900 Hz bandwidth (p=0.0028). This has implications for the application of silicon-embedded accelerometers for externally powered prosthesis control.

Amputation, Surgical↗

Acute renal failure requiring dialysis after allogeneic blood and marrow transplantation identifies very poor prognosis patients.

We examined the incidence, risk factors and associated mortality of acute renal failure requiring dialysis (Renal Bearman Grade [BG] 3) in a 3-year cohort of 97 consecutive allogeneic blood and marrow transplantation (alloBMT) patients. In all, 20 (21%) developed Renal BG3 (all died by day +132) and 77 (79%) developed renal insufficiency (Renal BG1-2). Renal BG3 was a contributing or primary cause of death in 18 (90%) patients who continued to require dialysis at time of death. The two Renal BG3 patients whose deaths were not related to renal failure died on day +103 of hemorrhage and day +132 of underlying disease. By univariate analysis, age, unrelated donor, veno-occlusive disease (VOD) and grade III-IV acute graft-versus-host disease with hepatic involvement were significantly associated with Renal BG3. The multivariate model of time to Renal BG3 determined only a prior diagnosis of severe acute GVHD (RR=4.1, 95% CI 1.6-10.3, P=0.003) and VOD (RR=9.1, 95% CI 3.5-23.7, P<0.001) as significant independent predictors. Renal BG3 is generally considered a conditioning regimen-related toxicity. This study demonstrates that Renal BG3 is most commonly a complication of hepatic co-morbidities after allogeneic blood and marrow transplantation and identifies patients with a very poor prognosis.

Acute Kidney Injury↗