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Biomedical subjects

A Sanchez

Publications and source records attributed to A Sanchez.

At least 109 records · Page 6Linked to original sources
PubMed1998

Variation in the glycoprotein and VP35 genes of Marburg virus strains.

Marburg virus, the prototype of the family Filoviridae, differs genetically, serologically, and morphologically from Ebola viruses. To better define the genetic variation within the species, VP35 and glycoprotein (GP) genes of representative human isolates from four known episodes of Marburg virus hemorrhagic fever were analyzed. The percentage nucleotide differences in the GP gene coding regions of Marburg viruses (0.1-21%) was nearly equal to the percentage amino acid changes (0-23%), while the percentage nucleotide differences in VP35 coding regions (0.3-20.9%) were higher than the percentage amino acid changes (0.9-6.1%), indicating a greater number of nonsynonymous changes occurring in the GP gene. The higher variation in the GP gene and the corresponding protein, especially those changes in the variable middle region of the GP, suggests that the variability may be the result of responses to natural host pressures. Analysis of the GP gene open reading frame shows a nonrandom distribution of nonsynonymous mutations that may indicate positive Darwinian selection is operating within the variable region. A heptad repeat region and an adjoining predicted fusion peptide are found in the C-terminal third of Marburg virus GPs, as has been previously shown for Ebola virus, and are similar to those found in transmembrane glycoproteins of retroviruses, paramyxoviruses, coronaviruses, and influenza viruses. Comparative analyses showed that there are two lineages within the Marburg virus species of filoviruses. The most recent isolate from Kenya (1987) represents a separate genetic lineage within the Marburg virus species (21-23% amino acid difference). However, this lineage likely does not represent a separate Marburg subtype, as the extent of divergence is less than that separating Ebola virus subtypes.

Amino Acid Sequence↗
PubMed1998

Use of clomiphene citrate in the treatment of men with high sperm chromatin stability.

OBJECTIVE: To determine whether improvements of the seminal vesicle function after a 5-day course with clomiphene citrate (CC) may reduce the prevalence of men with high sperm chromatin stability under conditions of sodium dodecyl sulfate (SDS)-ethylenediaminetetraacetic acid (EDTA). DESIGN: A prospective study. SETTING: Andrology laboratory at the Instituto de Investigaciones de la Altura, Universidad Peruana Cayetano Heredia, Lima, Peru. PATIENT(S): Forty-one male partners of infertile couples attending the andrology laboratory. INTERVENTION(S): Clomiphene citrate was administered orally twice a day. Men were treated with CC at 100 mg daily for 5 days. Blood and semen samples were collected before treatment and 24 hours after the last administration. MAIN OUTCOME MEASURE(S): Serum testosterone, seminal fructose, sperm motility, sperm chromatin stability after SDS and EDTA, and prevalence of high sperm chromatin stability. RESULT(S): The percentage of stable sperm after SDS-EDTA correlated inversely with the basal corrected concentration of seminal fructose (-1.77 +/- 0.89, beta +/- SE). High sperm chromatin stability was observed in 53.8% of the study population and in 66.7% of patients with hypofunction of the seminal vesicles. In those men whose seminal vesicle function improved after treatment with CC, the prevalence of high sperm chromatin stability was reduced from 67% to 25% (chi2 = 5.34). Logistic regression analysis showed that the higher the basal corrected seminal fructose levels and the higher the basal serum testosterone levels, the lower the probability of nonresponse of the sperm chromatin stability to treatment with CC (0.54 +/- 0.15, odds ratio +/- SE for corrected fructose; and 0.50 +/- 0.15, odds ratio +/- SE for serum testosterone). CONCLUSION(S): Hypofunction of the seminal vesicles was associated with high sperm chromatin stability, and this high sperm chromatin stability under SDS-EDTA conditions may be reduced by treatment with CC.

Adolescent↗
PubMed1998

A comparison of temperature measurements using three ear thermometers.

Following anecdotal and research-based reports of inaccuracies encountered with the use of ear thermometers in patient care settings, this two-phase study was designed to compare the accuracy of three ear thermometers when used in a multioperator environment. The within-subject variation (limits of agreement) in ear temperature measurements obtained with the three ear thermometers in a multioperator condition by using an oral temperature reference standard ranged from +/- 1.25 degrees F to 1.85 degrees F. In multioperator, multisubject patient care environments using a pulmonary artery catheter core temperature as the reference standard, the limits of agreement for ear temperature measurements obtained with the three different ear thermometers ranged from +/- 2.19 degrees F to 2.85 degrees F. These results suggest that there is substantial variation in ear measurements and raise questions about the use of ear thermometers when there are multiple personnel taking temperatures, as occurs in hospital-based clinical practice environments. Handedness of the operator, position of the patient, and the ear used for measurement did not produce clinically significant variability in ear temperature measurements.

Adult↗
PubMed1998

Immunization for Ebola virus infection.

Infection by Ebola virus causes rapidly progressive, often fatal, symptoms of fever, hemorrhage and hypotension. Previous attempts to elicit protective immunity for this disease have not met with success. We report here that protection against the lethal effects of Ebola virus can be achieved in an animal model by immunizing with plasmids encoding viral proteins. We analyzed immune responses to the viral nucleoprotein (NP) and the secreted or transmembrane forms of the glycoprotein (sGP or GP) and their ability to protect against infection in a guinea pig infection model analogous to the human disease. Protection was achieved and correlated with antibody titer and antigen-specific T-cell responses to sGP or GP. Immunity to Ebola virus can therefore be developed through genetic vaccination and may facilitate efforts to limit the spread of this disease.

Animals↗
PubMed1998

Morgagni's hernia resolved by laparoscopic surgery.

Morgagni's hernias represent between 2 and 4% of the whole of nontraumatic diaphragmatic hernias in the adult, and the treatment (even with minimal symptoms) is always with surgery. We present the case of a 50-year-old woman with an oppressive, postprandial discomfort in the right side of her thorax and with increased pain when supine. The x-ray examination indicated a large portion of transverse colon inside the thoracic cavity. Once the diagnosis of Morgagni's hernia had been obtained, she was scheduled for laparoscopic surgery to reduce the hernia and to reconstruct the defect of the diaphragm using a polypropylene mesh.

Female↗
PubMed1998

Intraspinal injection of adenosine agonists protect against L-NAME induced neuronal loss in the rat.

Intraspinal injection of the nonspecific inhibitor of nitric oxide synthase N-nitro-L-arginine methyl ester (L-NAME) results in a dose-dependent loss of neurons in the rat spinal cord. This effect is thought to result from a reduction in basal levels of nitric oxide (NO), thereby producing an ischemic reaction secondary to vasoconstriction and reduced spinal cord blood flow (SCBF). An important component of this ischemic reaction is the release of excitatory amino acids and the initiation of an excitotoxic cascade. In the present study, microinjections of adenosine A1 and A2 receptor agonists were made in the spinal cord to evaluate the neuroprotective effects of these drugs against neuronal loss produced by L-NAME. Animals were divided into six groups based on the composition of injected solutions: (a) L-NAME; (b) L-NAME + N6-cyclopentyladenosine (CPA, A1 agonist); (c) L-NAME + 5'-(N-cyclopropyl)-carboxamidoadenosine (CPCA, A2 agonist); (d) L-NAME + CPA + CPCA; (e) N-methyl D-aspartate (NMDA); and (f) NMDA + CPA. Injections of L-NAME or NMDA produced a unilateral loss of spinal neurons, a local inflammatory response, and darkly stained pyknotic nuclei surrounding the area of neuronal loss. CPA and CPCA significantly reduced the area of L-NAME-induced neuronal loss, and a synergistic effect was observed when ineffective doses of these agonists were co-injected with L-NAME. The excitotoxic effects of NMDA were not affected by CPA. The results have shown that A1 and A2 receptor agonists provide significant neuroprotection against L-NAME induced neuronal loss, presumably by inhibiting ischemia induced release of excitatory amino acids (A1 agonist), or by restoring SCBF secondary to vasodilation (A2 agonist). It is suggested by these results that the intraspinal injection of L-NAME is an effective model to study the pathological consequences of vasoconstriction, reduced SCBF, and ischemia secondary to decreased NO production in the rat spinal cord. Finally, the results provide support for the continued investigation of specific adenosine agonists as therapeutic agents directed against the ischemic and excitotoxic components of spinal injury.

Adenosine↗
PubMed1998

Filoviral haemorrhagic fevers.

Sensationalised accounts of wards of dying patients have fueled intense public fascination with filoviruses and highlighted the global threat of emerging and re-emerging infectious diseases. Filoviruses are the prototypical emerging pathogens: they cause a haemorrhagic disease of high case-fatality associated with explosive outbreaks due to person-to-person transmission, have no known treatment, occur unpredictably, and have an unknown reservoir. In truth, since their initial discovery in 1967, only a handful of filoviral outbreaks have occurred, mostly in remote locations. However, the documented occurrence of secondary cases in locations far from endemic areas validates the concern that filoviruses have the potential to cause unprecedented outbreaks in the future.

Filoviridae Infections↗
PubMed1998

Transpyloric enteral feeding in critically ill children.

BACKGROUND: Nutrition is important in childhood because the child has a lower energy reserve than the adult and a higher demand for calories because of ongoing growth. In this study, the utility of transpyloric enteral feeding (TEF) in critically ill children was evaluated. METHODS: A prospective, descriptive study was made in a pediatric intensive care unit of a tertiary pediatric center of 41 critically ill children, 30 after surgical procedures and 11 with nonsurgical illness, aged 8 days to 12 years, who received transpyloric enteral feeding with 8- or 10-Fr weighted feeding tubes. Analysis was made of tolerance and complications (vomiting, abdominal distension, excessive gastric residual, diarrhea, and pulmonary aspiration) of TEF. RESULTS: The mean duration of TEF was 19.5 +/- 26.8 days (range, 1-120 days). The administration of sedative agents or inotropic drugs did not alter toleration of TEF. Eight of 12 patients treated with continuous infusion of vecuronium tolerated TEF without complications. Eleven gastrointestinal complications occurred in 10 patients, abdominal distension and excessive gastric residual in 7 (17%), and diarrhea in 4 (9.7%). In 7 patients gastrointestinal complications improved, with decreasing use or transitory interruption of TEF, but in 4 patients (9.7%), TEF had to be withdrawn. Gastrointestinal complications were more frequent in postsurgical than in nonsurgical patients (p < 0.001). No patients suffered from pulmonary aspiration, and the incidence of pulmonary infection and hepatic dysfunction diminished during TEF. CONCLUSIONS: Transpyloric enteral feeding is a good method of nutritional support in critically ill children and can be used in patients treated with neuromuscular blocking agents. The frequency and severity of complications and the risks of pulmonary infection and hepatic dysfunction related to TEF are low.

Child↗
PubMed1998

Biochemical analysis of the secreted and virion glycoproteins of Ebola virus.

The glycoproteins expressed by a Zaire species of Ebola virus were analyzed for cleavage, oligomerization, and other structural properties to better define their functions. The 50- to 70-kDa secreted and 150-kDa virion/structural glycoproteins (SGP and GP, respectively), which share the 295 N-terminal residues, are cleaved near the N terminus by signalase. A second cleavage event, occurring in GP at a multibasic site (RRTRR downward arrow) that is likely mediated by furin, results in two glycoproteins (GP1 and GP2) linked by disulfide bonding. This furin cleavage site is present in the same position in the GPs of all Ebola viruses (R[R/K]X[R/K]R downward arrow), and one is predicted for Marburg viruses (R[R/K]KR downward arrow), although in a different location. Based on the results of cross-linking studies, we were able to determine that Ebola virion peplomers are composed of trimers of GP1-GP2 heterodimers and that aspects of their structure are similar to those of retroviruses, paramyxoviruses, and influenza viruses. We also determined that SGP is secreted from infected cells almost exclusively in the form of a homodimer that is joined by disulfide bonding.

Animals↗
PubMed1998

Helical CT versus CT arterial portography in the detection of hepatic metastasis of colorectal carcinoma.

OBJECTIVE: The study is a prospective comparison of helical CT with nonhelical CT arterial portography (CTAP) in the detection of liver metastases from colorectal carcinoma, using surgical and histologic findings as the gold standard. SUBJECTS AND METHODS: Thirty-five patients with colorectal carcinoma and suspected liver metastases were prospectively examined with helical CT and CTAP before surgery. In nine cases, surgery was not performed. In the remaining 26 patients, imaging results were correlated with surgical and pathologic findings. Three radiologists prospectively assessed metastatic involvement with both techniques. The results were compared with pathologic and surgical findings on a lesion-by-lesion basis. In a second phase, three radiologists not directly concerned in the design of the study independently assessed metastatic involvement of the liver as revealed on helical CT and CTAP on a segment-by-segment basis with a five-level scale of confidence. Results were analyzed by receiver operating characteristic methods. RESULTS: The results of the histologic study disclosed 50 metastatic lesions. Helical CT had a sensitivity of 76% (38/50) and a positive predictive value of 90%. CTAP had a sensitivity of 74% (37/50) and positive predictive value of 69%. Receiver operating characteristic analysis revealed a greater area under the curve (Az index), 0.96, for helical CT than for CTAP (0.86). Differences were statistically significant (p < .001). CONCLUSION: Helical CT is superior to nonhelical CTAP in the detection of hepatic metastases from colorectal carcinoma.

Adult↗
PubMed1997

A system for functional analysis of Ebola virus glycoprotein.

Ebola virus causes hemorrhagic fever in humans and nonhuman primates, resulting in mortality rates of up to 90%. Studies of this virus have been hampered by its extraordinary pathogenicity, which requires biosafety level 4 containment. To circumvent this problem, we developed a novel complementation system for functional analysis of Ebola virus glycoproteins. It relies on a recombinant vesicular stomatitis virus (VSV) that contains the green fluorescent protein gene instead of the receptor-binding G protein gene (VSVDeltaG*). Herein we show that Ebola Reston virus glycoprotein (ResGP) is efficiently incorporated into VSV particles. This recombinant VSV with integrated ResGP (VSVDeltaG*-ResGP) infected primate cells more efficiently than any of the other mammalian or avian cells examined, in a manner consistent with the host range tropism of Ebola virus, whereas VSVDeltaG* complemented with VSV G protein (VSVDeltaG*-G) efficiently infected the majority of the cells tested. We also tested the utility of this system for investigating the cellular receptors for Ebola virus. Chemical modification of cells to alter their surface proteins markedly reduced their susceptibility to VSVDeltaG*-ResGP but not to VSVDeltaG*-G. These findings suggest that cell surface glycoproteins with N-linked oligosaccharide chains contribute to the entry of Ebola viruses, presumably acting as a specific receptor and/or cofactor for virus entry. Thus, our VSV system should be useful for investigating the functions of glycoproteins from highly pathogenic viruses or those incapable of being cultured in vitro.

Animals↗
PubMed1997

The role of cysteinyl residues in the activity of bacterial elongation factor Ts, a guanosine nucleotide dissociation protein.

The modification of E.coli elongation factor Ts (EF-Ts) by NEM and other sulfhydryl reagents inactivates the protein's ability to bind EF-Tu.GDP and to catalyze GDP exchange. The reactive residue was found to be Cys-22. Replacement of Cys-22 by Ser or Gly only partially impairs the binding or catalytic properties of EF-Ts while it completely protects EF-Ts from the inactivation by NEM. Cys-22 of EF-Ts is not located at the EF-Ts.EF-Tu interface, yet it can be modified only when EF-Ts is not bound to EF-Tu. These results support the proposal that the conformation change around Cys-22 in the amino terminus of EF-Ts rather than Cys-22 itself is essential for binding EF-Tu. Apparently, modification of Cys-22 by NEM disrupts the conformation change and inactivates EF-Ts. The return of EF-Ts to its native conformation may provide the driving force for the rate-determining step in the catalytic cycle, the dissociation of EF-Ts from EF-Tu.GNP.

Amino Acid Substitution↗
PubMed1997

Completion of the Lassa fever virus sequence and identification of a RING finger open reading frame at the L RNA 5' End.

Lassa (LAS) fever virus is a highly pathogenic arenavirus with large (L) and small (S) RNA genomic segments. The 5' end of the LAS L segment is described here, thereby completing the sequence of the most virulent arenavirus analyzed to date. In keeping with the ambisense gene structure of the arenaviruses, the LAS L RNA encodes a 250-kDa protein and an 11-kDa protein in opposite senses with respect to each other. The 11-kDa protein, defined previously in arenaviruses lymphocytic choriomeningitis (LCM), Tacaribe (TAC), and Pichinde (PIC), contains a RING type of zinc-binding structure. Expression of the 11-kDa protein in LAS virus-infected cells has been confirmed by binding to peptide-specific antibody.

Amino Acid Sequence↗
PubMed1997

The repeat expansion detection method in the analysis of diseases with CAG/CTG repeat expansion: usefulness and limitations.

The repeat expansion detection (RED) method was described to detect expansions of trinucleotide repeats of unknown chromosomal location. We have improved the RED method by the use of 8-mer oligonucleotides and assessed its usefulness in 30 samples from patients with spinocerebellar ataxia type 1 (SCA1), Huntington's disease (HD), and Machado Joseph's disease (MJD), for which the number of CAG/CTG repeats was determined by sequencing. There was a good correlation between the number of repeats detected by sequencing and those identified by RED. However, in 17% of samples, the RED gave additional fragments for ligation products of different size than the CAG/CTG repeat expansion detected in the sample by sequencing. The same was observed in a group of control subjects (n = 78) without known clinical abnormalities in which products of more than 40 repeats were detected in 27% of them, indicating that CAG/CTG repeat expansions are common in the general population. Wether this corresponds to unidentified loci with expansions deserves further investigation.

DNA↗
PubMed1997

Capillary electrophoresis.

Capillary electrophoresis is a rapid and versatile electrophoretic technique that has found several applications in the field of peptide synthesis. The high resolution and charge-based separation capabilities make it very complementary to RP-HPLC. In addition, both are essentially quantitative, which makes them ideal as purity assessment techniques. Capillary electrophoresis can be used to monitor any charge-related changes in synthetic peptides such as incomplete deprotection or chemical modifications. Capillary electrophoresis can also be very sensitive to changes in the shape or size of a peptide. However, its greatest value for the characterization of synthetic peptides is when it is used in combination with other analytical techniques such as mass spectrometry, HPLC, and amino acid analysis.

Amino Acid Sequence↗
PubMed1997

Isolation of genomic DNA from milk samples by using Chelex resin.

A rapid procedure for isolating genomic DNA from milk samples has been devised, based on the use of Chelex resin. By using this protocol, genomic DNA was extracted from milk samples from 15 cows and 15 goats. The suitability of these DNA preparations as a template for performing the polymerase chain reaction (PCR) was tested by amplifying three different loci of the bovine genome: exon 4 of the kappa-casein gene and the INRA5 and INRA23 microsatellites, together with two others: exon 19 of the alpha s1-casein gene and exon 2 and part of intron 2 of the DRB gene of the caprine genome. No amplification products could be obtained from any sampless at 30 cycles. In contrast, at 45 cycles the number of amplified samples ranged from 86 to 100% and at 65 cycles all the DNA targets were amplified, indicating that the number of cycles was a critical factor to be optimized for obtaining the desired PCR target. These results suggest that this method may be a useful tool for analysing genetic polymorphism at the DNA level by PCR and relating it to milk composition and other traits of economic interest.

Animals↗
PubMed1997

Galanin and cholecystokinin in cultured magnocellular neurons isolated from adult rat supraoptic nuclei: a correlative light and scanning electron microscopical study.

Cultured magnocellular neurons, isolated from adult rat supraoptic nuclei, were characterized by immunocytochemistry, using the avidin-biotin-peroxidase complex and antisera to vasopressin, oxytocin, galanin and cholecystokinin. Light microscope examination of the immunostained cultures revealed the presence of vasopressin- and oxytocin-like immunoreactivity, as well as neurons containing either galanin- or cholecystokinin-like immunoreactivity. In contrast, no significant galanin- or cholecystokinin-like immunoreactivity could be observed in freshly dispersed cells. Correlative scanning electron microscopical observations in the secondary electron imaging mode revealed that the stained neurons appeared significantly brighter than the unstained structures. Complementary observations with toad brain sections (preoptic area), immunostained for galanin, led to the same result. Considering previous results, it is suggested that the presence of galanin- and cholecystokinin-like immunoreactivity in the cultured neurons and its virtual absence in freshly dispersed cells is indicating a participation of these peptides in the regenerative processes taking place during culture. It is further concluded that the avidin-biotin-peroxidase method is suitable for correlative light and scanning electron microscopical studies of smooth surfaces and cultured cells.

Animals↗