Search PubMed⌕ Search

Biomedical subjects

D Martinez

Publications and source records attributed to D Martinez.

At least 73 records · Page 4Linked to original sources

MR features of developing periventricular white matter in preterm infants: evidence of glial cell migration.

PURPOSE: MR imaging of the brain is increasingly used in the investigation of the newborn, but little information is available on the normal appearance of the developing brain. We scanned a series of newborn infants in an attempt to define the normal appearance of developing periventricular white matter and to assess how pathologic conditions may modify this appearance. METHODS: Sixty-eight newborn infants, median postmenstrual age (PMA) 34 weeks (range, 24 to 42 weeks), were subdivided into two groups: group A (n = 33), which included those with normal clinical and sonographic examinations, and group B (n = 35), which contained those with evidence of neuroabnormality detected prior to the MR study, either clinically or by cerebral sonography. Images were acquired in two planes on a 1.5-T imager using turbo spin-echo pulse sequences. RESULTS: Symmetric periventricular bands of reduced signal intensity were noted in the frontal periventricular white matter on T2-weighted images in 98% of group A infants and in 97% of group B infants. The number of bands was inversely related to PMA. The reduction in number of bands with increasing PMA was delayed in group B infants. CONCLUSION: The uniform appearance of periventricular bands in a population of healthy infants and their relationship to the infants' maturity is consistent with the results of previous histologic studies. These studies demonstrate the presence of migrating glial cells within the periventricular white matter of infants beyond 20 weeks' gestation, when neuronal migration to the cortex is complete. We postulate that the bands seen on T2-weighted images represent groups of migrating glial cells, providing a further marker of cerebral maturation.

Brain↗

Detection of genomic polymorphisms among isolates of the intracellular bacterium Cowdria ruminantium by random amplified polymorphic DNA and Southern blotting.

Sixteen primers were successfully used in a RAPD assay to generate reproducible fingerprints for six isolates of Cowdria ruminantium, a tick-transmitted rickettsia of ruminants. Distinction between stocks was possible by using one or at most two primers. Two stocks were very similar although originating from widely distant geographical regions. A genetic distance tree was constructed by analysing 108 fragments in pairwise comparison between stocks. Three amplification fragments probed with C. ruminantium genomic DNA determined a restriction fragment length polymorphism which allowed the distinction between stocks except for the two stocks that had similar RAPD patterns. The potential of RAPD to determine the extent of genetic diversity of C. ruminantium and to develop probes or PCR primers for diagnostic purposes is discussed.

Blotting, Southern↗

The prognostic significance of intratumoral natural killer cells in patients with colorectal carcinoma.

BACKGROUND: Natural killer (NK) cells have a spontaneous cytotoxic capacity-against tumor cells. These cells represent a small proportion of human colon carcinoma-infiltrating lymphocytes. Their prognostic significance in these tumors has yet to be determined. METHODS: One hundred and fifty-seven patients who each had a colectomy for large bowel adenocarcinoma were studied. No patient received adjuvant therapy. Immunohistochemical stains were performed for NK cells using the monoclonal antibody CD57. The number of NK cells was counted using a MICRON image analyzer. The total area studied for each tumor was 1 cm2. In this area, 50 intratumoral fields of 0.173 mm2 were selected. The degree of NK infiltration was classified as little (< 50 NK cells), moderate (50-150 NK cells), and extensive (> 150 NK cells). The Kaplan-Meier method was used to obtain survival figures. Multivariate analyses were performed using the Cox regression model. RESULTS: At 5 years, patients with little and moderate NK infiltration showed significantly shorter survival rates (overall and disease free survival) than those with extensive infiltration (P < 0.01). Three significant factors affecting survival were selected in a stepwise fashion in increasing order as follows: TNM stage, NK infiltration, and lymphocytic infiltration. Patients with TNM Stage III disease and extensive NK infiltration showed significantly longer survival rates than those with little or moderate infiltration (P < 0.001). In these patients, multivariate analysis using the Cox regression model identified two significant variables: number of involved lymph nodes and NK cells infiltration. CONCLUSIONS: In patients with colorectal carcinoma, an extensive intratumoral infiltration of NK cells is associated with a favorable tumor outcome. Intratumoral infiltration of NK cells can be used as a variable with prognostic value, especially in patients with TNM Stage III disease.

Adenocarcinoma↗

Gastroschisis: a radiological and clinical review.

A retrospective clinico-radiological review was undertaken of 66 consecutive cases of gastroschisis managed at our institution between August 1982 and February 1993. The condition's morbidity and mortality were reviewed, as were its radiological features and their impact upon management. All patients underwent surgery in the first 24 h of life, and the overall survival rate was 92 %. The finding of bowel atresia at operation was associated with a particularly poor outcome, with only two out of five infants surviving. A minority of infants developed serious complications including necrotizing enterocolitis, short-bowel syndrome, persistent small-bowel dysfunction and cholestatic jaundice. Investigation by plain films, contrast studies and ultrasound examinations was necessary and helpful in these patients. Plain film radiography commonly revealed bowel-wall thickening and luminal dilatation, frequently accompanied by generalised abdominal distension. Small-bowel enema was considered to be superior to the conventional follow-through in distinguishing mechanical from functional obstruction in infants with persistent bowel dilatation.

Abdominal Muscles↗

Development of an in vitro cloning method for Cowdria ruminantium.

Cowdria ruminantium is a tick-borne rickettsia which causes severe disease in ruminants. All studies with C. ruminantium reported so far were carried out with stocks consisting of infective blood collected from reacting animals or from the same stocks propagated in vitro. Cloned isolates are needed to conduct studies on immune response of the host, on genetic diversity of the parasite, and on mechanisms of attenuation and the development of vaccines. A method of cloning based on the particular chlamydia life cycle of Cowdria was developed. Instead of cloning extracellular elementary bodies, it appeared more convenient to clone endothelial cells infected by one morula resulting from the infection of the cell by one elementary body of Cowdria. Two hundred and sixteen clones were obtained by limiting dilution of infected cells. The method was experimentally validated by comparing randomly amplified polymorphic DNA fingerprints from individual clones obtained from endothelial cell cultures coinfected with two different stocks of C. ruminantium.

Animals↗

Analysis of T-cell responses in cattle immunized against heartwater by vaccination with killed elementary bodies of Cowdria ruminantium.

Cattle were successfully immunized against heartwater with a lysate of Cowdria ruminantium formulated in Freund's adjuvant. Vaccinated animals proved fully resistant to virulent challenge 3 and 10 months after vaccination. For the first time a helper T lymphocyte response to Cowdria antigens was observed and characterized. Cowdria-specific T-cell lines generated from vaccinated animals by in vitro restimulation with Cowdria lysates are 95 to 100% CD4+, are MHC class II restricted, and produce gamma interferon. They proliferate in response to autologous monocytes infected with live Cowdria but not in response to uninfected monocytes. These T-cell lines will facilitate the search for Cowdria antigens that are immunogenic for T cells and will therefore be of relevance in the development of a subunit vaccine against the disease.

Animals↗

Development and evaluation of a recombinant antigen, monoclonal antibody-based competitive ELISA for heartwater serodiagnosis.

Cowdria ruminantium is the etiologic agent of heartwater, a tick-transmitted foreign animal disease with considerable potential for entrance into the USA. A competitive enzyme-linked immunosorbent assay (cELISA) was developed to detect serologic responses to C. ruminantium infection. The cELISA utilized a recombinant form of the C. ruminantium major antigenic protein (MAP-1) as the antigen and an anti-MAP-1 monoclonal antibody as the competing indicator reagent. Experimental antisera to C. ruminantium and a wide variety of related ehrlichial organisms were used to evaluate cELISA reactivity. Only sera against C. ruminantium, Ehrlichia canis, E. chaffeensis, and a recently discovered cervine ehrlichia-like organism reacted positively in the cELISA. Specificity of the cELISA was > or = 99.5% in a survey of 1,774 southeastern US and Puerto Rican slaughter cattle sera but was only 85% in a group of 79 hunter-killed white-tailed deer (Odocoileus virginianus) from the southeastern USA. Reference true-positive and cELISA false-positive sera were further analyzed by end point titrations using the cELISA and by indirect fluorescent antibody (IFA) tests for reactivity with C. ruminantium, E. canis, and E. chaffeensis antigens. True heartwater-positive sera were significantly more reactive using the cELISA and C. ruminantium IFA procedures (P < 0.05), whereas false-positive sera were significantly more reactive with the antigens used in the E. chaffeensis IFA procedure (P < 0.05). A group of sera from 210 field-origin ruminants residing on known or potentially heartwater-endemic Caribbean islands revealed a substantial (12.4%) prevalence of cELISA-positive specimens. The cELISA is a relatively specific serodiagnostic test for heartwater in cattle and could be used to monitor for possible introduction of the disease into the USA. The cELISA may also be an excellent tool for monitoring the success of an ongoing Caribbean Amblyomma tick eradication program designed to eliminate the biological vector responsible for the perpetuation and spread of this dangerous foreign animal disease.

Animals↗

Tricompartmental kinetics of the organophosphorous pesticide dimethoate.

We present a study on the distribution and persistence of the organophosphorous pesticide, dimethoate, in the Wistar rat. Dimethoate's levels in blood and tissues were determined using gas chromatography with a NPD detector. The toxicokinetic profile was estimated by non-linear regression using the software program PCNONLIN, which concluded that a tricompartmental model best described dimethoate's behavior in the rat body.

Adipose Tissue↗

Comparative efficacy of Freund's and Montanide ISA50 adjuvants for the immunisation of goats against heartwater with inactivated Cowdria ruminantium.

Two vaccines, based on inactivated elementary bodies of Cowdria ruminantium, one formulated in Montanide ISA50, the other in Freund's adjuvant, were compared in goats. Administered twice subcutaneously with an interval of 81 days, both protected three out of five goats against a very severe challenge, lethal for all 14 control goats, 3.5 months after the second injection. Both vaccines elicited similar antibody levels. The protection afforded by the Montanide ISA50 vaccine was tested 15 and 17 months after the second injection of the vaccine. Three out of six and five out of six goats, respectively, survived a challenge which killed all four control goats used on each occasion. Antibodies were still detectable in the immunised goats. The level of protection appears to be influenced by the dose of virulent C. ruminantium used for the challenge. As any stock of C. ruminantium can be incorporated in order to cover the antigenic repertoire of the organism, this kind of inactivated vaccine can now be tested in the field.

Adjuvants, Immunologic↗

Stent embolization during intracoronary stenting.

Nine patients with systemic embolization complicating intracoronary stenting (incidence 1.8%) are described. Although this rare complication was not associated with any clinical sequelae, great care should be taken to prevent this possibility, especially in patients with unfavorable anatomic characteristics, including lesions in the left circumflex artery, at bend points, and calcified lesions.

Aged↗

Recombinant expression and use in serology of a specific fragment from the Cowdria ruminantium MAP1 protein.

The major antigenic protein (MAP1) of Cowdria ruminantium was screened for immunogenic regions by expression of overlapping recombinant DNA clones of the gene encoding the MAP1 protein. Two regions, designated MAP1-A and MAP1-B, were recognized by all antisera to 9 different isolates of C. ruminantium. MAP1-A contained one or more epitopes responsible for false-positive reactions with Ehrlichia antisera in several serological tests for cowdriosis. Cross-reactivity with MAP1-B was limited to antisera to Ehrlichia chaffeensis and Ehrlichia canis. Antisera to Ehrlichia species that infect ruminants (E. bovis, E. ovina, and E. phagocytophila) did not recognize MAP1-B. The sensitivity of an indirect ELISA based on MAP1-B was found to be excellent, since all sera from animals experimentally infected with C. ruminantium (64 out of 64) reacted with MAP1-B. Validation of this ELISA was carried out with field sera obtained from sheep raised in heartwater-free areas in Zimbabwe and from several Caribbean islands. Only 9 out of 111 samples from Zimbabwe, and 1 out of 58 samples from the Caribbean islands, which were considered to be false positives by immunoblot or indirect ELISA, reacted with MAP1-B. Thus, the ELISA based on MAP1-B is at present the most specific and sensitive serological test for cowdriosis.

Animals↗

An amino acid sequence coded by the exon 2 of the BoLA DRB3 gene associated with a BoLA class I specificity constitutes a likely genetic marker of resistance to dermatophilosis in Brahman zebu cattle of Martinique (FWI).

One hundred and twenty-seven Brahman cattle from several locations in Martinique (FWI), reared under different environmental conditions, were followed over three years and checked for clinical signs of dermatophilosis. To confirm that these animals had been in contact with the pathogen Dermatophilus congolensis, their sera were tested by ELISA. On the basis of this epidemiological study, 12 animals were classified as resistant (seropositive without clinical signs), belonging to herds in which the prevalence of the disease ranged from 25 to nearly 98%. Eighteen animals classified as highly susceptible displayed severe characteristic skin lesions. These 30 selected animals were typed for class I antigens of the major histocompatibility complex (MHC). MHC class II genes were analyzed using the polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) techniques, on the exon 2 of the bovine leucocyte antigen (BoLA) DRB3 gene. Several alleles were found, according to patterns provided by the restriction enzymes used: Fnu 4HI, Dpn II, Hae III, and Rsa I. A particular sequence "EIAY" at amino acid positions 66/67/74/78 located in the antigen recognition sites (ARS) was found in the 12 animals classified as resistant, and 10 of them displayed also class I BoLA-A8 specificity. On the other hand, only 3 out of the 18 susceptible animals showed simultaneously the BoLA-DRB3 "EIAY" sequence and BoLA-A8 specificity. Interestingly, a serine residue at position 30 of the ARS was found in 8 of the susceptible animals and was completely absent from all resistant animals. Furthermore, in a same animal, the serine at position 30 and the EIAY sequence were never found simultaneously on the same haplotype. These results show a strong correlation between the resistant character to dermatophilosis and the association of MHC haplotypes: the BoLA-A8 specificity and the BoLA-DRB3 "EIAY" sequence at ARS positions 66/67/74/78 with the lack of serine in position 30. To confirm these results, family segregation studies are in progress and some interesting observations have been obtained.

Actinomycetales Infections↗

Recombinant bovine interferon gamma inhibits the growth of Cowdria ruminantium but fails to induce major histocompatibility complex class II following infection of endothelial cells.

Recombinant bovine IFN gamma is a potent inhibitor of Cowdria ruminantium growth in vitro irrespective of the rickettsial stock, or the origin of the endothelial cells. These results suggest an important role for IFN gamma in protective immune responses against C. ruminantium infections. Here we also show that IFN gamma can induce the expression of MHC class II molecules on the surface of endothelial cells. However, treatment of endothelial cells with IFN gamma following infection with Cowdria fails to induce MHC class II expression. The implications of this pathogen-specific effect on class II expression by endothelial cells with regard to its recognition by the host immune system are discussed.

Animals↗

Fetal gallstones.

Three male fetuses examined sonographically in late gestation to assess growth and well-being were found to have evidence of gallbladder cholelithiasis. Postnatally, all three were asymptomatic but they had variable sonographic findings. In two, the abnormalities noted sonographically had resolved spontaneously by 6 weeks of age, but the third patient had a persistent gallbladder "calculus" at 6 months. All three boys, now aged 9 months to 4 years, remain well, with no evidence of biliary tract disease. Sonographic evidence of fetal gallbladder cholelithiasis may be found in late gestation, is more common among boys, and frequently is associated with benign spontaneous resolution.

Adult↗

Open thoracotomy procedural competency: validity study of teaching and assessment modalities.

STUDY OBJECTIVES: To determine (1) reliability and validity estimates of three modalities used to assess open thoracotomy procedural competency and (2) the effect of computer practice on procedural performance as measured by the three assessment modalities. METHODS: An experimental, sequential assessment design with volunteer examinees completing all three assessment modalities (paper, computer, pig model) was implemented at the animal support facilities of a university medical school with an affiliated emergency medicine residency program. Level of physician training (student, resident, faculty) and type of computer practice (thoracotomy, cricothyrotomy) were independent variables. Procedural competency scores were determined for each modality; scores were defined in terms of performance time and performance accuracy for three thoracotomy procedures (opening the chest, pericardiotomy, and aortic cross-clamping). RESULTS: Thoracotomy performance on the pig reliably discriminated among examinees known to differ in level of training. However, computer simulation performance did not significantly differ among examinees with different levels of training. Computer simulation practice significantly improved later performance on the computer assessment (P < .05) but not on the pig assessment. The greatest predictor of procedural competency (time and accuracy) on the pig assessment was the ability to sequentially order procedural steps. CONCLUSION: This study establishes the pig model as superior to the paper and computer models as the criterion standard for open thoracotomy assessment. Psychometric properties support the pig model as the most reliable and valid model yet described for assessing thoracotomy procedural competency. Computer simulation practice using visual images (complex anatomy) and the sequential ordering of procedural steps through paper modeling show promise for teaching and assessment of prerequisite skills required to develop psychomotor procedural competency.

Animals↗