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Biomedical subjects

E Guzman

Publications and source records attributed to E Guzman.

At least 19 recordsLinked to original sources

A rapid and accurate assay for assessing the cytotoxicity of viral proteins.

A fluorescence-based assay is presented for measuring the cytoxicity of viral proteins added exogenously to cells. The assay is based on the use of two fluorescent dyes, calcein-AM and ethidium homodimer (EtD-1) to specifically stain living and dead cells respectively and employs fluorescence activated cells sorting (FACS) to achieve a rapid and accurate measurement of the cytotoxic capacity of a potential viral toxin. The assay has been developed using the group B homologue (ADRV-NSP4) of the NSP4 enterotoxin encoded by Group A rotaviruses but should be applicable to assaying any viral protein exhibiting cytotoxic activity.

Cytotoxicity Tests, Immunologic↗

Molecular characterization of the rotavirus NSP4 enterotoxin homologue from group B rotavirus.

The RNA segment (Gene 10) from a human group B rotavirus which encodes the homologue of the rotavirus enterotoxin (NSP4) has been cloned and sequenced. The gene is of the same length (751 nucleotides) as its better-characterized group A rotavirus counterpart but shows minimal homology (approximately 10%) to it at the primary sequence level. Despite this low level of sequence homology, secondary structure predictions for the group B protein (ADRV-NSP4) showed a close similarity of structural features with the group A protein. Full-length ADRV-NSP4 was expressed in Escherichia coli with an amino terminal 6xHis tag that was used to purify it to homogeneity. The cytotoxicity of the purified protein was examined in a rapid dye-uptake assay that assesses membrane permeability and was found to be comparable to its group A counterpart.

Amino Acid Sequence↗

[Multidisciplinary approach of the obese child to the dietary residency of "Clairs Vallons"].

The pediatric obesity has become a real problem for the public health. One estimates that about 16% of the Belgian pediatric population and up to 33% of the Americans are concerned by this problematic. 70% of the teenagers will remain obese once adult if no treatment is proposed during the childhood. Because of that evolution, some journalists wrote: "that the old continent would be able to catch up with the new world in the next ten years". The malnutrition is not however the only factor at the origin of the obesity. The sedentary lifestyle (lack of exercise, TV, Internet, video games) the domestic organization, the "various emotional stress "are to be blamed. It is without taking into consideration the paradox of our consumption society that while extolling the cult of the slim, young and dynamic body, etc., pushes us to consume more, encouraging in some social and cultural surroundings to go for the immediate pleasure to the detriment of the knowledge - understanding - of our own body. Which places the obese child in an existential paradox. If on top of it there is a domestic predisposition to the plumpness, the kilograms in excess are threatening the unsecured child and new sufferings stand out on the horizon : relational unrests, isolation, social dismissal, reduction of the esteem of selfesteem as well as lack of confidence, less freedom, depression, etc. Not only are they victims of mockeries, aggressiveness and exclusion, the children put their health in danger. On those children we can notice an increase of the impact of cardiovascular pathologies, diabetes, cancers of the intestines, etc. In "Clairs Vallons" we put the hypothesis that the children and the teenagers who come here in custody could suffer from a lack of presence as well as people listening to them and that therefore to would search for comfort in eating. We consider that all interventions based solely on the interdiction of the symptom have no result, causes more suffering and a displacement of the symptom. The therapeutic work to the Dietary Residency of "Clairs Vallons" consists therefore in a global approach of the child within a multidisciplinary team (pediatrician, dietitian, psychologist, physiotherapist, social assistant, orthofenist, educator and teacher) all united around the concept "to eat happy".

Child↗

HGT-DB: a database of putative horizontally transferred genes in prokaryotic complete genomes.

The Horizontal Gene Transfer DataBase (HGT-DB) is a genomic database that includes statistical parameters such as G+C content, codon and amino-acid usage, as well as information about which genes deviate in these parameters for prokaryotic complete genomes. Under the hypothesis that genes from distantly related species have different nucleotide compositions, these deviated genes may have been acquired by horizontal gene transfer. The current version of the database contains 88 bacterial and archaeal complete genomes, including multiple chromosomes and strains. For each genome, the database provides statistical parameters for all the genes, as well as averages and standard deviations of G+C content, codon usage, relative synonymous codon usage and amino-acid content. It also provides information about correspondence analyses of the codon usage, plus lists of extraneous group of genes in terms of G+C content and lists of putatively acquired genes. With this information, researchers can explore the G+C content and codon usage of a gene when they find incongruities in sequence-based phylogenetic trees. A search engine that allows searches for gene names or keywords for a specific organism is also available. HGT-DB is freely accessible at http://www.fut.es/~debb/HGT.

Amino Acids↗

Mad dogs, Englishmen and apoptosis: the role of cell death in UV-induced skin cancer.

Apoptosis plays a critical role in the development and progression of ultraviolet-induced skin cancers. In particular, Fas and Fas ligand (FasL) interactions are known to control the development of "sunburn cells" or apoptotic keratinocytes in the UV-exposed epidermis. In the absence of functional Fas/FasL signaling, UV-induced apoptosis is diminished and mutations rapidly accumulate. UV-induced suppression of host immunity, a process regulating skin cancer outgrowth, is also controlled through Fas/FasL interactions. Other death receptors, such as the receptor for tumor necrosis factor, may also contribute to UV-induced carcinogenesis and progression. Understanding the involvement of cell death in cancers caused by exposure to sunlight may provide novel approaches for prevention and therapy of these ever-increasing malignancies.

Animals↗

The role of the cytoskeleton in capacitative calcium entry in myenteric glia.

Capacitative calcium entry (CCE) is the process by which intracellular calcium is replenished from the external milieu upon depletion of intracellular stores. CCE is thought to participate in chemotaxis, proliferation and cell signalling. A physical interaction between intracellular stores and the plasma membrane is postulated to regulate CCE. We hypothesized that cytoskeletal disruption alters this interaction, inhibiting CCE in enteric glia. Cultured myenteric glia from neonatal guinea-pigs were treated with cytochalasin D (10 micro mol L-1), a microfilament disrupting agent, nocodazole (20 micro mol L-1), a microtubule disrupting agent, or vehicle (dimethyl sulphoxide). Intracellular calcium changes were measured using fura-2 microfluorimetry. To evaluate the rate of cation re-entry, barium was substituted for calcium because barium is not sequestered internally. Cytochalasin D-treated glia had diminished CCE responses (57 +/- 3 nmol L-1) compared with controls (97 +/- 7 nmol L-1) as did nocodazole-treated glia (30 +/- 2 nmol L-1) vs controls (77 +/- 6 nmol L-1). The proportion of cells demonstrating CCE abolition was greater in the cytochalasin (50 +/- 8%) and nocodazole-treated (89 +/- 2%) groups compared with controls (21 +/- 2%, 40 +/- 9%, respectively). Cytochalasin D and nocodazole treatment diminished the rate of cation re-entry based on diminished barium entry in treated vs control cells. From this study, we conclude that disruption of cytoskeletal elements diminishes calcium influx essential to calcium store repletion in myenteric glia.

Animals↗

Design and implementation of a measuring campaign to model pesticide impacts in an Ecuadorian watershed.

The first important issue in pesticide assessment for a watershed is the availability of soil and water quality information in the form of charts, maps or records. However, developing countries generally do not have enough historical data. Thus, sampling programs are crucial for the success of the evaluation, although in developing countries they always represent a controversial task between limited available budget and a precise assessment. This paper shows the steps taken to generate soil and water data to be used in a pesticide assessment project for a 34,000 ha watershed in Ecuador, South America. Sampling campaigns are still being run, so the methodology and the first results are shown here.

Ecuador↗

Chromatin-dependent cooperativity between constitutive and inducible activation domains in CREB.

The cyclic AMP (cAMP)-responsive factor CREB induces target gene expression via constitutive (Q2) and inducible (KID, for kinase-inducible domain) activation domains that function synergistically in response to cellular signals. KID stimulates transcription via a phospho (Ser133)-dependent interaction with the coactivator paralogs CREB binding protein and p300, whereas Q2 recruits the TFIID complex via a direct association with hTAF(II)130. Here we investigate the mechanism underlying cooperativity between the Q2 domain and KID in CREB by in vitro transcription assay with naked DNA and chromatin templates containing the cAMP-responsive somatostatin promoter. The Q2 domain was highly active on a naked DNA template, and Ser133 phosphorylation had no additional effect on transcriptional initiation in crude extracts. Q2 activity was repressed on a chromatin template, however, and this repression was relieved by the phospho (Ser133) KID-dependent recruitment of p300 histone acetyltransferase activity to the promoter. In chromatin immunoprecipitation assays of NIH 3T3 cells, cAMP-dependent recruitment of p300 to the somatostatin promoter stimulated acetylation of histone H4. Correspondingly, overexpression of hTAFII130 potentiated CREB activity in cells exposed to cAMP, but had no effect on reporter gene expression in unstimulated cells. We propose that cooperativity between the KID and Q2 domains proceeds via a chromatin-dependent mechanism in which recruitment of p300 facilitates subsequent interaction of CREB with TFIID.

Animals↗

Use of cotton rats for preclinical evaluation of measles vaccines.

The continued prevalence and medical impact of measles worldwide has created interest in the development of new generations of measles vaccines. Monkeys can be used for preclinical testing of these vaccines. However, a more practical and less expensive animal model is highly desirable, particularly for initial vaccine development and evaluation. Cotton rats have been shown to support the replication of different strains of measles virus (MV), and thus may be useful for these purposes. To test this concept, the immunogenicity and protective efficacy of two standard (Moraten and trivalent measles, mumps, rubella) and four experimental (two recombinant ALVAC, one ISCOM subunit and live attenuated Edmonston-Zagreb) MV vaccines were evaluated in naïve cotton rats, and cotton rats with passively acquired MV-specific neutralizing serum antibodies. All of the test vaccines were immunogenic and protected naíve animals from pulmonary infection and viral dissemination. However, under the conditions utilized, only the Edmonston-Zagreb vaccine provided such protection to animals with significant levels of passively acquired MV-specific neutralizing antibodies. The results of these tests and the potential of using cotton rats as an animal model for preliminary testing of MV vaccines are discussed.

Animals↗

Resolution of ST-segment elevation after streptokinase therapy in anterior versus inferior wall myocardial infarction.

BACKGROUND: Resolution of ST-segment elevation is the best bedside predictor of myocardial reperfusion. HYPOTHESIS: This study was conducted to examine the resolution of ST-segment elevation after streptokinase therapy in anterior versus inferior acute myocardial infarction (MI) and to corroborate it with echocardiographic and coronary angiographic data. METHODS: The study population consisted of 70 patients, 35 each in the anterior and inferior MI groups. The electrocardiograms (ECGs) were recorded before, on completion of, and on Days 1 and 2 post streptokinase therapy. The resolution of ST segment determined from post-streptokinase ECGs was compared between the two groups and correlated with echocardiographic and coronary angiographic data. RESULTS: On completion of and on Day 1 post streptokinase therapy, ST-segment resolution in both groups was not significantly different. On Day 2 post streptokinase therapy, resolution of the ST segment per lead was significantly lower in anterior than that in inferior MI (61 +/- 21% anterior vs. 77 +/- 21% inferior, p 0.003). The number of patients with akinesis of infarct-related ventricular wall was significantly higher (17 anterior vs. 7 inferior, p 0.02), and left ventricular ejection fraction was significantly lower in anterior MI (39 +/- 7% anterior vs. 48 +/- 8% inferior, p < 0.01). There was no significant difference in coronary angiographic data. One patient in each group demonstrated normal coronary arteries. CONCLUSIONS: The resolution of ST-segment elevation on the completion of and on Day 1 post streptokinase therapy was comparable between anterior and inferior MI. The significantly less frequent resolution of ST-segment elevation in anterior MI on Day 2 post streptokinase could be due to more akinesis, larger infarct size, and worse systolic function rather than due to failure to open the infarct-related vessel.

Coronary Angiography↗

Nephrogenic adenofibroma in a young child.

Nephrogenic adenofibroma is a benign renal tumor in children and young adults described by Hennigar and Beckwith in 1992. Seven cases have been described, and we report the first case in an 11-month-old child, in good health, revealed by a macroscopic hematuria. Nephrogenic adenofibroma is an unusual tumor, which was difficult to distinguish from nephroblastoma and mesoblastic nephroma. Beckwith makes a distinction between this principal differential diagnosis in child renal tumors based upon morphologic and immunohistochemical patterns. In our observation, the diagnosis remained difficult and needed several reviews of our case. Beckwith proposed the final diagnosis: nephrogenic adenofibroma with stromal predominance. The prognosis is excellent and no treatment is indicated. A FISH analysis of the tumor cells found a trisomy 11. Trisomy 11 has been reported in mesoblastic nephroma as the most frequent chromosomal abnormality. This finding in tumor cells provides an argument for excluding the diagnosis of nephroblastoma but can not clarify the difference between nephrogenic adenofibroma and mesoblastic nephroma.

Adenofibroma↗

Use of cotton rats to evaluate the efficacy of antivirals in treatment of measles virus infections.

No practical animal models for the testing of chemotherapeutic or biologic agents identified in cell culture assays as being active against measles virus (MV) are currently available. Cotton rats may serve this purpose. To evaluate this possibility, 5-ethynyl-1-beta-D-ribofuranosylimidazole-4-carboxamide (EICAR) and poly(acrylamidomethyl propanesulfonate) (PAMPS), two compounds that have been reported to inhibit MV in vitro, and ribavirin, an established antiviral drug with MV-inhibitory activity, were evaluated for their antiviral activities against MV and respiratory syncytial virus (RSV) in tissue culture and in hispid cotton rats. A single administration of PAMPS markedly inhibited pulmonary RSV or MV replication (>3 log(10) reduction in pulmonary titer compared to that for controls), but only if this compound was administered intranasally at about the time of virus inoculation. Both EICAR and ribavirin exhibited therapeutic activity against RSV and MV in cotton rats when they were administered parenterally. However, both of these compounds were less effective against MV. On the basis of the pulmonary virus titers on day 4 after virus inoculation, the minimal efficacious dose of EICAR against MV (120 mg/kg of body weight/day when delivered intraperitoneally twice daily) appeared to be three times lower against this virus than that of ribavirin delivered at a similar dose (i.e., 360 mg/kg/day). These findings correlated with those obtained in vitro. The data obtained suggest that cotton rats may indeed be useful for the initial evaluation of the activities of antiviral agents against MV.

Animals↗

Neutrophil F-actin and myosin but not microtubules functionally regulate transepithelial migration induced by interleukin 8 across a cultured intestinal epithelial monolayer.

The role of the polymorphonuclear leukocyte (PMN) cytoskeleton during the transmigration across colonic epithelial cells is not very well understood. In order to study the role of different components of the PMN cytoskeleton during transepithelial migration across a colonic epithelial cell monolayer (T84), PMN were preincubated with drugs affecting either the actin cytoskeleton (cytochalasin B, iota toxin of Clostridium perfringens, and phalloidin) or the microtubules (colchicine and taxol). The role of PMN myosin during transepithelial migration was investigated using the inhibitor 2,3-butanedione monoxime (BDM) and DC3B toxin. PMN intracellular Ca2+, during neutrophil adhesion and translocation across the epithelium, was assessed by the Ca2+ chelator 1, 2bis-(2-aminophenoxy)-ethane-N,N,N', N'-tetra-acetic acid tetrakis (acetoxymethyl) ester (BAPTA-AM). Transmigration of PMN was initiated by applying either interleukin-8 or formyl-met-leu-phe (fMLP). While colchicine and taxol preexposure did not influence PMN transepithelial migration, treatment with cytochalasin B, iota toxin, phalloidin, BDM, DC3B toxin and BAPTA-AM greatly diminished migration of PMN across T84 monolayers. Similarly, cell-cell contacts established between PMN and epithelial cells during the transmigration were diminished after treatment of PMN with iota toxin or cytochalasin B. These data show that the neutrophil actin cytokeleton and myosin, but not the microtubules, evoke a Ca2+ -dependent motility that facilitates migration across the colonic epithelial barrier.

ADP Ribose Transferases↗

[Fibrous splenic hamartoma: imaging features].

Splenoma or splenic hamartoma is a rare primary splenic tumor most often incidentally discovered. The authors report the case of a splenic hamartoma, developed in an asymptomatic man and imaged by US, CT and MRI. This tumor which measured 7 cm in diameter, was hypoechoic with posterior hardening of the ultrasound beam, and not much vascularized as demonstrated by Duplex and color Doppler examination. The tumor MR imaging showed hyperintensity on T1-weighted and hypointensity on T2-weighted. The final diagnosis was not established until the histologic examination of the splenectomy specimen was performed. Histopathologically, the tumor corresponded to a fibrous type of splenic hamartoma. The atypical imaging features which led to splenectomy were probably related to the fibrous component of the hamartoma.

Adult↗

Gallbladder rupture during laparoscopic cholecystectomy: does it have an effect on postoperative morbidity?

Gallbladder rupture during laparoscopic cholecystectomy is a common event that may lead to increased postoperative morbidity. To evaluate this event, we reviewed 300 cases of laparoscopic cholecystectomy. Duration of surgery and hospitalization, postoperative symptoms, wound infection, and late complications were analyzed by comparing two groups of patients, one without gallbladder rupture (A) and one with rupture (B). Gallbladder rupture was found in 40 cases (13.9%). Duration of surgery averaged 81 min for group A and 96.5 min for group B. Postoperative symptoms in the first 24 hours were present in approximately 10% of patients in both groups. Within the first 24 hours, 92.3% of patients in group A were discharged compared with 85% in group B. One patient (0.4%) in group A developed wound infection compared with 2 patients (5%) in group B (p = 0.05). To date, no patients have developed late abdominal complications associated with the procedure. Although this was a retrospective and uncontrolled study, gallbladder rupture during laparoscopic cholecystectomy was found to be associated with increased wound infections. No other significant effects on postoperative morbidity were detected.

Adult↗

The nucleotide sequence of Saccharomyces cerevisiae chromosome V.

Here we report the sequence of 569,202 base pairs of Saccharomyces cerevisiae chromosome V. Analysis of the sequence revealed a centromere, two telomeres and 271 open reading frames (ORFs) plus 13 tRNAs and four small nuclear RNAs. There are two Tyl transposable elements, each of which contains an ORF (included in the count of 271). Of the ORFs, 78 (29%) are new, 81 (30%) have potential homologues in the public databases, and 112 (41%) are previously characterized yeast genes.

Base Sequence↗