Search PubMed⌕ Search

Biomedical subjects

J Silva

Publications and source records attributed to J Silva.

At least 145 records · Page 8Linked to original sources
PubMed1995

IgG subtype is correlated with efficiency of passive protection and effector function of anti-herpes simplex virus glycoprotein D monoclonal antibodies.

IgG subclasses differ in their effector functions in a variety of in vitro assays. To assess the effect of antibody subclass differences on in vivo protective efficacy against herpes simplex virus (HSV), a series of subclass switch mutants was made from an anti-HSV glycoprotein D monoclonal antibody. Purified antibody was examined for the ability to protect against HSV-2 challenge in mice. IgG2a was found to be more effective than IgG1. This correlated both with activity in antibody-dependent cell-mediated cytotoxicity and with efficiency of complement-mediated neutralization. These data suggest that optimization of passive immunization against HSV requires consideration of antibody subclass.

Animals↗
PubMed1995

Comparison of arbitrarily primed PCR with restriction endonuclease and immunoblot analyses for typing Clostridium difficile isolates.

Arbitrarily primed PCR (AP-PCR) was used to genotype 26 clinical isolates of Clostridium difficile previously analyzed by immunoblotting (IB) and 20 isolates typed by restriction endonuclease analysis (REA) with HindIII. Two levels of differentiation were achieved with the AP-PCR approach by use of two different arbitrary primers. With the 19-mer arbitrary primer T-7 (first level of differentiation), a good correlation was found between IB and AP-PCR typing. Twenty isolates grouped into six IB types were separated into seven major AP-PCR types. These seven AP-PCR groups were further discriminated into 12 subtypes after genotyping with the arbitrary primer PG-05 (second level of differentiation). The remaining six isolates, all of different IB types, showed a unique and distinct DNA banding pattern with both of the arbitrary primers, T-7 and PG-05. Twenty isolates representing 20 REA types from 15 REA groups were resolved into 13 AP-PCR DNA profiles with the arbitrary primer T-7. A good correlation was found at this level of differentiation between the major REA groups, Y and M, and AP-PCR typing. While AP-PCR with this primer failed to differentiate isolates in REA groups J, G, R, and B, AP-PCR with PG-05 resolved these four isolates into four distinct AP-PCR types. In addition, one of three M strains and one of four Y strains displayed a slightly different DNA banding pattern by AP-PCR (with PG-05) from that of the other strains in the group. We conclude that AP-PCR is a rapid and sensitive method which not only complements other typing schemes but also may be a substitute and prove to be especially suited for immediate epidemiological tracking of nosocomial infections due to C. difficile.

Bacterial Typing Techniques↗
PubMed1995

Genomic fingerprinting of epidemic and endemic strains of Stenotrophomonas maltophilia (formerly Xanthomonas maltophilia) by arbitrarily primed PCR.

Arbitrarily primed PCR (AP-PCR) was used to type 64 clinical isolates of Stenotrophomonas maltophilia from 60 patients and the hands of one nurse. Forty-seven different patterns were observed, most patients having isolates with unique genomic fingerprints. A single pattern, however, was obtained from six of eight patients involved in an intensive care nursery outbreak, confirming the suspected nosocomial transmission of this microorganism. This strain was also found in four other patients hospitalized at the same time but in different units. AP-PCR typing results had a good correlation with the 49 patterns obtained when the isolates were typed by contour-clamped homogeneous electric field gel electrophoresis. Although AP-PCR is slightly less discriminatory than contour-clamped homogeneous electric field gel electrophoresis, it offers several advantages and should be considered as a practical option for molecular typing during investigations of outbreaks.

Bacterial Typing Techniques↗
PubMed1995

Apparent outbreaks of Clostridium difficile-associated diarrhea in horses in a veterinary medical teaching hospital.

Intestinal colonization with toxigenic strains of Clostridium difficile was documented in 9 of 10 horses with acute onset diarrhea in a veterinary medical teaching hospital, whereas a similar isolate was detected in only 1 of 23 other horses without diarrhea in the hospital. One horse with diarrhea was infected simultaneously with both C. difficile and Salmonella krefeld. Clostridium difficile was detected by fecal culture on selective medium, confirmed with a latex particle agglutination test, and identified as toxigenic by polymerase chain reaction amplification of toxin A and toxin B gene sequences. Using an arbitrarily-primed polymerase chain reaction, 6 distinct C. difficile isolates were detected in the feces of the 9 affected horses at the time of the outbreak of diarrhea.

Animals↗
PubMed1995

[Hospital prevalence of kidney failure. Consequences and reflections for the planning of nephrology services].

A prospective prevalence study of Renal Failure (RF) in inhospital patients (creatinine > = 1.7 mg/dl) was carried out during March 1994, in two hospitals covering well defined and mutually exclusive populations. Cases were selected by screening all urea and creatinine blood tests performed in both laboratories, and registered in an individual form for daily follow-up of their nephrologic outcome. We registered 242 RF cases among 3525 patients admitted (6.8%), with an A.R.F. prevalence of 5.2%, 46% of all patients had a serum creatinine > 3 mg/dl and 71% were older than 65 years. In 55% RF was acquired inside the hospital. The most frequent cause of A.R.F. was pre-renal failure with 37%, followed by 32% of ischemic acute tubular necrosis and 13% toxic ATN. Nephrology was consulted in only 29% of all hospital RF cases. Only 17% of the RF patients were submitted to dialysis procedures, overall mortality was 31%, and 30% had normal renal function at discharge. Our results provide a data base to rethink the organization, staffing and role of nephrology departments inside general hospitals.

Female↗
PubMed1995

The Scenario-Based Engineering Process (SEP): a user-centered approach for the development of health care systems.

The Scenario-based Engineering Process (SEP) is a user-focused methodology for large and complex system design. This process supports new application development from requirements analysis with domain models to component selection, design and modification, implementation, integration, and archival placement. It is built upon object-oriented methodologies, domain modeling strategies, and scenario-based techniques to provide an analysis process for mapping application requirements to available components. We are using SEP in the health care applications that we are developing. The process has already achieved success in the manufacturing and military domains and is being adopted by many organizations. SEP should prove viable in any domain containing scenarios that can be decomposed into tasks.

Artificial Intelligence↗
PubMed1994

Cytosolic-free calcium elevation in Trypanosoma cruzi is required for cell invasion.

To replicate, the trypomastigote form of Trypanosoma cruzi must invade host cells. Since a role for Ca2+ in the process of cell invasion by several intracellular parasites has been postulated, changes in the intracellular Ca2+ concentration in T. cruzi trypomastigotes and in tissue culture L6E9 myoblasts during their interaction were studied at the single cell level using digital imaging fluorescence microscopy or in cell suspensions by fluorescence spectrophotometry. An increase in cytosolic Ca2+ in T. cruzi trypomastigotes was detected at the single cell level after association of the parasites with the myoblasts. Ca2+ mobilization in the host cells was also detected upon contact with trypomastigotes either at the single cell level or in cells grown in coverslips and exposed to suspensions of trypomastigotes. Pretreatment of the parasites with the Ca2+ chelators quin 2 (50 microM) or bis-(o-aminophenoxy)-ethane-N,N,N',N'-tetraacetic acid (BAPTA, 50 microM) decreased the trypomastigotes' association to myoblasts by approximately 40 and 63%, respectively, thus indicating that an increase in intracellular Ca2+ concentration in the parasites is required for cell invasion in addition to Ca2+ mobilization in the host cells.

Animals↗
PubMed1994

Specific detection of Clostridium difficile toxin A gene sequences in clinical isolates.

The polymerase chain reaction (PCR) was used to specifically detect toxin A gene sequences of Clostridium difficile in DNA isolated from human faeces. A set of oligonucleotide primers derived from the non-repetitive region of the toxin A gene was developed to amplify a 634-bp DNA fragment. All 28 cytotoxic strains of C. difficile, previously characterized by a toxin B-PCR assay, were positive for the presence of toxin A gene sequences. No amplification products were obtained from DNAs extracted from non-toxigenic strains, strains of C. sordellii, or C. bifermentans. In addition, amplification of DNA extracted from C. difficile 8864, a strain which does not produce toxin A, resulted in multiple bands which probed negative for toxin A gene sequences. DNAs extracted from nine stool specimens which were positive for toxin B by the cytotoxicity assay and by the toxin B-PCR assay were also positive in this assay. Toxin A gene sequences were detected in DNAs obtained from 4/11 stool specimens which were negative by the toxin B cytotoxicity assay. These four specimens were from patients who had a history of relapses due to C. difficile-associated colitis, and whose stools had previously been found to be positive by the toxin B-PCR test despite no detectable toxin B in the specimens. These data indicate a comparable degree of clinical sensitivity between these two toxin-gene PCR-based assays. This rapid, sensitive and specific assay may be useful not only in the diagnosis of C. difficile infections, but also in molecular studies of the toxin A gene in C. difficile strains.

Bacterial Proteins↗
PubMed1994

Health care professional workstation: software system construction using DSSA scenario-based engineering process.

This paper describes a new method for the evolutionary determination of user requirements and system specifications called scenario-based engineering process (SEP). Health care professional workstations are critical components of large scale health care system architectures. We suggest that domain-specific software architectures (DSSAs) be used to specify standard interfaces and protocols for reusable software components throughout those architectures, including workstations. We encourage the use of engineering principles and abstraction mechanisms. Engineering principles are flexible guidelines, adaptable to particular situations. Abstraction mechanisms are simplifications for management of complexity. We recommend object-oriented design principles, graphical structural specifications, and formal components' behavioral specifications. We give an ambulatory care scenario and associated models to demonstrate SEP. The scenario uses health care terminology and gives patients' and health care providers' system views. Our goal is to have a threefold benefit. (i) Scenario view abstractions provide consistent interdisciplinary communications. (ii) Hierarchical object-oriented structures provide useful abstractions for reuse, understandability, and long term evolution. (iii) SEP and health care DSSA integration into computer aided software engineering (CASE) environments. These environments should support rapid construction and certification of individualized systems, from reuse libraries.

Computer Systems↗
PubMed1994

Endoscopic bile duct stone removal prior to laparoscopic cholecystectomy.

Of 1049 patients referred for laparoscopic cholecystectomy (LC) for symptomatic gallstone disease, 67 (6%) had clinical, biochemical, or echographic findings suggesting common bile duct stones. Patients in this group were studied preoperatively with endoscopic retrograde cholangiopancreatography (ERCP). In 26 patients (39%), the diagnosis was confirmed. In 12 other cases (18%), the macroscopic finding of a stripped or bleeding papilla without common bile duct stones suggested stone migration. ERCP in the remaining 29 patients (43%) was normal. Thirty-four endoscopic sphincterotomies (ES) were performed, 26 for common bile duct stones and 8 for cystic lithiasis or gallbladder microlithiasis. In the entire group of patients with choledocholithiasis, stone removal was possible. All 67 patients underwent laparoscopic cholecystectomy on an average of 2.8 days following the endoscopic procedure. Twenty-one patients (31%) had acute cholecystitis, and 5 had chronic scleroatrophic cholecystitis. Five (7.5%) of the 67 patients were converted to an open procedure. In 10 cases (16%), the cystic diameter was larger than an 8-mm M-L clip, which made necessary the use of endoligature or extra clips. No complications or deaths resulted from ERCP or ES. Two of the 62 patients (3.2%) who underwent LC had to be reoperated on, 1 because of a right subphrenic collection, and the other because of bilious ascites. No common bile duct lesions or deaths resulted in the analyzed group. The average hospitalization time, with the exception of those patients converted or reoperated on, was 8 days.

Adolescent↗
PubMed1994

Genotyping of Clostridium difficile isolates.

Arbitrarily primed polymerase chain reaction (AP-PCR) was used to genotype Clostridium difficile isolates from various sources. Four major molecular types were identified among strains from the American Type Culture Collection previously typed by serogroup and from isolates from patients at the University of California, Davis Medical Center, from a patient at a Utah institution, and from the environment. These groups contained subgroups that displayed, in addition to the common group bands, at least one unique band. Two strains isolated from patients at our institution had the same DNA banding patterns. These patients were hospitalized during the same period, raising the possibility of cross-infection through hospital contact or another common source. These results suggest that this AP-PCR approach will be useful in epidemiologic studies of C. difficile infections.

Base Sequence↗
PubMed1994

Fecal shedding of Clostridium difficile in dogs: a period prevalence survey in a veterinary medical teaching hospital.

The goal of this study was to determine the fecal prevalence of Clostridium difficile in dogs who were patients at a veterinary medical teaching hospital. Stool specimens collected from 152 dogs (in- and outpatients) were analyzed for the presence of C. difficile. An additional 42 stool specimens were collected and examined from dogs recently housed at local animal shelters. Following culture on selective medium, C. difficile was identified by a latex agglutination test, and the presence of the toxin A and B genes was determined individually by polymerase chain reaction. Clostridium difficile was isolated from the feces of 28 of the veterinary hospital patients (18.4%); isolates from 14 of these patients (50.0%) were toxigenic. Diarrhea was a clinical finding in 5 (35.7%) of the dogs carrying toxigenic isolates of C. difficile, whereas diarrhea was noted in only 2 of 14 dogs (14.3%) shedding nontoxigenic isolates. Three of 14 dogs (21.4%) shedding toxigenic isolates of C. difficile were receiving antibiotics at the time of stool collection, whereas 5 of 14 dogs (37.5%) shedding nontoxigenic strains of C. difficile were receiving antibiotics. The carriage rate of C. difficile was significantly higher for animals categorized as inpatients of the veterinary hospital. The carriage rate also provided evidence for an increased risk for fecal shedding with increasing age. Clostridium difficile was not isolated from any of the 42 dogs recently housed at local animal shelters. This study confirms the presence of toxigenic C. difficile in dogs at a veterinary teaching hospital.(ABSTRACT TRUNCATED AT 250 WORDS)

Age Factors↗
PubMed1994

[Laparoscopic cholecystectomy makes difficult the analysis of gallbladder mucosa. Morphometric study].

Laparoscopic gallbladder surgery damages gallbladder mucosa, impeding an adequate histological examination. We studied gallbladder samples coming from 56 conventional and 44 laparoscopic cholecystectomies performed in patients with chronic cholecystitis without signs of acute inflammation. The length of each examined sample and the length of the portion where superficial epithelia was not found was measured. No differences in sample length was found between both types of cholecystectomy. However; the segment with superficial mucosa was significantly shorter in the samples coming from laparoscopic surgery. Seventeen samples did not contain mucosa at all and 14 of these (82%) came from laparoscopic procedures. Eighty six percent of samples coming from laparoscopic surgery were considered inadequate for histological study compared to 36% of samples coming from conventional cholecystectomy. It is concluded that laparoscopic cholecystectomy hampers the study of gallbladder mucosa and may have an impact on the diagnosis of neoplastic gallbladder lesions.

Cholecystectomy↗
PubMed1994

[Laparoscopic cholecystectomy. 1203 cases with ultraselective cholangiography].

We present our experience at the University of Chile Hospital with 1203 cases of laparoscopic cholecistectomy between April 1991 and July 1993. In only 36 patients ultraselective cholangiography was performed. In 88 cases an endoscopic cholangiography was performed prior to surgery because of suspicion of choledocolithiasis. In 31 of them choledocolithiasis was demonstrated, and was successfully treated by papilotomy in 30 of them. In 5.8% of cases coversion to open surgery was necessary: 1.4% in chronic cholecystitis, 13% in acute cholecystitis and 39% in cases with fibrosis and esclerosis of gallbladder with or without biliary digestive fistula. The intraoperative cholangiography has been advocated as a manner to prevent the common duct injury. In spite of the small number of cases operated on with cholangiography, we haven't seen serious injuries of biliary tract; a carefully disection, as far away as possible of the common duct, is the best way to prevent iatrogenic lesions. Postoperative morbidity was present in 15 cases, 8 of them required a new operation. Two patients died.

Adolescent↗