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D Martinez

Publications and source records attributed to D Martinez.

At least 55 records · Page 3Linked to original sources

Effect of isolation techniques, in vitro culture and IFNgamma treatment on the constitutive expression of MHC Class I and Class II molecules on goat neutrophils.

Previous studies on the ability of human neutrophils to synthesize cytokines and express MHC Class I and inducible Class II molecules have suggested a possible role of these cells as accessory or antigen presenting cells (APC). There is no information available to date concerning this aspect in ruminants. Therefore, as the first step of these investigations, we have studied the cell surface expression of MHC molecules on goat neutrophils. We show that goat neutrophils can be distinguished from eosinophils with monoclonal antibody (MoAb) ILA-24 which recognizes cattle monocytes and neutrophils. Goat neutrophils constitutively express MHC Class II molecules. However, cell surface expression of MHC Class I and Class II molecules is dramatically reduced on neutrophils purified by density gradient centrifugation in comparison to neutrophils obtained from whole blood after lysis of erythrocytes. Also, the level of expression of MHC Class I antigens is seasonal and donor-dependent and rapidly decreases after in vitro culture despite negligible necrosis and apoptosis of neutrophils. Although treatment with IFNgamma partially prevents the loss of MHC Class I molecules on neutrophils, it fails to induce MHC Class II antigens. Implications of these results for further studies on the potential role of neutrophils as APC are discussed.

Animals↗

Immune responses to Cowdria ruminantium infections.

Understanding the basis of protective immunity to Cowdria ruminantium will facilitate the development of an effective subunit vaccine against heartwater in ruminants and contribute to a better definition of protective immune mechanisms to obligate intracellular pathogens in general. Until recently, immunological studies of heartwater in ruminants concentrated solely on antibody responses. Since 1995, the mechanisms underlying cell-mediated immunity of heartwater have been analysed. Progress achieved in these areas is discussed here by Philippe Totté and colleagues, with special emphasis on ruminants, the natural hosts of C. ruminantium.

Animals↗

Kinetics and mechanism of St I modification by peroxyl radicals.

St I is a toxin present in the Caribbean Sea anemone Stichodactyla helianthus which is highly hemolytic in the nanomolar concentration range. Exposure of the toxin to free radicals produced in the pyrolysis of 2,2'-azobis(2-amidinopropane) hydrochloride leads to a progressive loss of hemolytic activity. This loss of hemolytic activity is accompanied by extensive modification of tryptophan residues. On the average, three tryptophan residues are modified by each inactivated toxin. The loss of hemolytic activity of St I takes place without significant changes in the protein structure, as evidenced by the similarity of the fluorescence and CD spectra of native and modified proteins. Also, the native and modified ensembles present a similar resistance to their denaturation by guanidinium chloride. The hemolytic behavior and the performance of the toxin at the single-channel level when incorporated to black lipid membranes suggest that the modified ensemble can be considered as composed of inactive toxins and active toxins whose behavior is similar to that of the native proteins. These results, together with the lack of induction time in the activity loss, suggest that the fall of hemolytic activity takes place by an all-or-nothing inactivation mechanism in which the molecules become inactive when a critical amino acid residue is modified.

Amino Acids↗

Developing protein documentaries and other multimedia presentations for molecular biology.

Computer-based multimedia technology for distance learning and research has come of age--the price point is acceptable, domain experts using off-the-shelf software can prepare compelling materials, and the material can be efficiently delivered via the Internet to a large audience. While not presenting any new scientific results, this paper outlines experiences with a variety of commercial and free software tools and the associated protocols we have used to prepare protein documentaries and other multimedia presentations relevant to molecular biology. A protein documentary is defined here as a description of the relationship between structure and function in a single protein or in a related family of proteins. A description using text and images which is further enhanced by the use of sound and interactive graphics. Examples of documentaries prepared to describe cAMP dependent protein kinase, the founding structural member of the protein kinase family for which there is now over 40 structures can be found at http://franklin.burnham-inst.org/rcsb. A variety of other prototype multimedia presentations for molecular biology described in this paper can be found at http://fraklin.burnham-inst.org.

Computer-Assisted Instruction↗

Rostral and orbital prefrontal cortex dysfunction in the manic state of bipolar disorder.

OBJECTIVE: This study investigated prefrontal cortex function in the manic state of bipolar disorder. METHOD: High-sensitivity [15O]H2O positron emission tomography and a word generation activation paradigm were used to study regional cerebral blood flow in five manic and six euthymic individuals with bipolar disorder and in five healthy individuals. RESULTS: Decreased right rostral and orbital prefrontal cortex activation during word generation and decreased orbitofrontal activity during rest were associated with mania. CONCLUSIONS: The data support the presence of rostral and orbital prefrontal dysfunction in primary mania. These findings, when seen in the context of the human brain lesion and the behavioral neuroanatomic literatures, may help to explain some of the neurobehavioral abnormalities characteristic of the manic state.

Adult↗

Tick-borne rickettiosis in Guadeloupe, the French West Indies: isolation of Rickettsia africae from Amblyomma variegatum ticks and serosurvey in humans, cattle, and goats.

Twenty-seven rickettsiae were isolated and/or detected from 100 Amblyomma variegatum ticks collected on Guadeloupe in the French West Indies. In this study, the polymerase chain reaction procedure appeared to be more sensitive in detecting rickettsiae in ticks than the shell-vial technique. Sequencing a portion of the outer membrane protein A-encoding gene showed that these rickettsiae appeared to be identical to Rickettsia africae, a member of the spotted fever group rickettsiae recently described as an agent of African tick-bite fever occurring in sub-Sahelian Africa. A high seroprevalence of antibodies to R. africae was demonstrated among mammals, particularly humans, cattle, and goats. These results and a recently reported case of an infection due to R. africae on Guadeloupe demonstrate that R. africae is present on this island. Although this disease has been underdiagnosed there, it may be frequent and may exist on other Caribbean islands where A. variegatum has propagated dramatically over recent years.

Adolescent↗

HIV/AIDS risks among Native American drug users: key findings from focus group interviews and implications for intervention strategies.

A multisite study funded through the National Institute on Drug Abuse and the Office of Research on Minority Health was conducted in 1996 to determine the HIV/AIDS prevention needs of Native American out-of-treatment drug users. In an effort to recommend directions for HIV/AIDS prevention programming, one component of this study entailed conducting a series of focus groups at each of four sites: Anchorage, Alaska; Denver, Colorado; Flagstaff, Arizona; and Tucson, Arizona. While some site differences were noted, several consistent thematic findings were revealed across all locations. Specifically, focus group members strongly recommended directly involving key members of the Native American community in conducting outreach and intervention activities, involving Native people as the sources of information, and utilizing local and tribally relevant forms of delivering the message. Other consistent themes included getting messages to smaller communities to prevent the potential "annihilation" of tribes, educating youth, and linking alcohol prevention education to HIV/AIDS education. Findings from this study support the idea that future HIV/AIDS prevention programs must take into account subgroup and individual level differences among Native American drug users.

Acquired Immunodeficiency Syndrome↗

Inhibitory effect of Cowdria ruminantium on the expression of MHC class I and class II molecules on bovine endothelial cells.

Bovine endothelial cells constitutively express MHC class I molecules, whereas MHC class II molecules can be induced by interferon gamma (IFN gamma) treatment. Endothelial cells may play a role as antigen presenting cells (APC), but their role in the immune response against Cowdria ruminantium is unknown. We have studied the effect of C. ruminantium infection on the expression of MHC class I and class II molecules on the surface of bovine endothelial cells. Bovine brain endothelial cells (BBEC) from the microvasculature and bovine umbilical endothelial cells (BUEC) from the macrovasculature were cultured in vitro, infected with different concentration of Cowdria, and treated or not with IFN gamma. We observed that Cowdria induced a reduction of MHC molecules expression on the surface of BUEC and BBEC. This inhibitory effect of Cowdria on MHC molecules could affect the capacity of endothelial cells to act as efficient antigen-presenting cells.

Animals↗

Validation and comparison of three enzyme-linked immunosorbent assays for the detection of antibodies to Cowdria ruminantium infection.

Serological tests for Cowdria ruminantium infection have been hampered by low specificity. Here, an indirect ELISA based on purified antigen, a competitive ELISA using a recombinant major antigenic protein (MAP-1) and an indirect ELISA based on the MAP-1B region of the recombinant MAP-1 were compared. The tests were validated using 3000 sera of ruminants from 14 islands of the Lesser Antilles as well as sequential serum samples from 10 cattle, 17 goats and 10 sheep vaccinated with inactivated C. ruminantium in ISA 50 adjuvant and from 14 goats infected with a virulent culture supernatant. All tests detected significantly higher percentages of positives on Antigua, Guadeloupe and Marie-Galante, where C. ruminantium had been isolated before. Overall specificity calculated with sera from the other 11 heartwater-free islands was 98.1%, 98.5%, and 99.4% for the ELISA based on crude antigen, recombinant MAP-1 and MAP-1B, respectively. Sensitivities observed with sequential serum samples were similar for all tests. Tests based on recombinant antigens, especially the MAP-1B, showed improved specificity, suggesting their use for epidemiological studies in regions where the distribution of cowdriosis is unknown. In addition, the competitive ELISA is useful for studies in wildlife for which species-specific conjugates do not exist.

Animals↗

Inhibition of MHC class I and class II cell surface expression on bovine endothelial cells upon infection with Cowdria ruminantium.

Endothelial cells constitute a main target for Cowdria ruminantium (CR) and can potentially play a role as antigen presenting cells (APC). Therefore, we measured, in vitro, the effect of CR infections on the expression of MHC class I and class II molecules on bovine umbilical endothelial cells (BUEC) and on bovine brain endothelial cells (BBEC). A dramatic inhibition of the expression of IFNgamma induced MHC class II molecules was observed on BUEC and to a lesser extent on BBEC upon CR infection. This inhibitory effect was also observed on constitutively expressed MHC class I molecules. Part of the reduction of cell surface MHC molecules could be ascribed to their accumulation in intracellular compartments pinpointing a disruption in the transit of these molecules to the surface of the cells. The exact mechanisms of inhibition are not yet known but, as opposed to what is described in other models, the involvement of prostaglandin E2 can be excluded. The results obtained in this study show that endothelial cells have a decreased capacity to express both MHC class I and class II molecules on their surface upon CR infection, thus favouring the escape of this pathogen from the host immune system.

Animals↗

Case report: cerebellar hemi-hypoplasia.

Cerebellar hypoplasia is a prominent feature of fetal brain pathology. The lesion is rarely isolated or asymmetric. Various aetiological factors have been proposed. The frequency of cerebellar hypoplasia as a congenital defect in humans is unknown. A single case is described of unilateral cerebellar hypoplasia with intact vermis that was detected on prenatal ultrasound examination. Changes in the appearance of the abnormality over the course of the pregnancy raised the possibility of a disruptive vascular aetiology with destruction of normally formed structures. Prenatal ultrasound, post-delivery cranial ultrasound, and MRI images are presented to support the hypothesis.

Adult↗

Modification of sticholysin II hemolytic activity by free radicals.

Sticholysin II is a highly hemolytic toxin present in the caribbean sea anemone Stichodactyla helianthus. Pre-incubation of St II with 2,2'-azobis(2-amidinopropane), a source of peroxyl radicals in air saturated solution, readily reduces its hemolytic activity. Analysis of the amino acids present in the protein after its modification shows that only tryptophan groups are significantly modified by the free radicals. According to this, the loss of hemolytic activity correlates with the loss of the protein intrinsic fluorescence. The results indicate that, at high toxin concentrations, nearly a tryptophan residue and 0.2 toxin molecules are inactivated by each radical introduced into the system. Association of St II to multilamellar liposomes (egg yolk phosphatidyl choline:sphingomyelin 1:1) increases the toxin intrinsic fluorescence, indicating a more hydrophobic average environment of the five tryptophan groups of the protein. In agreement with this, incorporation of St II to the liposomes reduces the rate of fluorescence loss during its modification by free radicals, particularly at long incubation times. These results are explained in terms of two populations of tryptophans that are quenched at different rates by acrylamide and whose rates of inactivation by free radicals are also different.

Acrylamide↗

Characterization of variable immunodominant antigens of Cowdria ruminantium by ELISA and immunoblots.

Cross-immunization experiments have revealed a significant antigenic diversity of the isolate of Cowdria ruminantium which needs to be characterized for the development of vaccines. We identified polymorphic immunodominant antigens by ELISA and immunoblot. Using serum from a goat immune to the Gardel stock of Cowdria (isolated in Guadeloupe) adsorbed on antigen of the Senegal stock of this pathogen, distinct serogroups were revealed by ELISA among six isolates from different geographical origins. Furthermore, a goat serum directed against the Senegal stock and adsorbed on Gardel antigens was shown to be specific for the Senegal stock, thus confirming the existence of serotypes in Cowdria. The Major Antigenic Protein 1 (MAP1) of Cowdria was shown to have variable antigenic determinants. Also in a group of variable proteins ranging from 23 to 29 kDa, one antigen of 26-27 kDa had a determinant specific for the Gardel isolate. These polymorphic antigens may be relevant components of Cowdria ruminantium for a vaccine as the sera revealing these antigens originated from a goal surviving a lethal challenge. However, the presence of T-cell epitopes and the ability of the these antigens to confer protection to ruminants remain to be investigated. The production of a rabbit antiserum against this group of polypeptides will be of great use for their purification and for the screening of expression libraries.

Animals↗

Reporting requirements for skeletal digital radiography: comparison of soft-copy and hard-copy presentation.

PURPOSE: To assess diagnostic performance and reader preference when reporting results from digital hard-copy and two soft-copy formats of skeletal digital radiography. MATERIALS AND METHODS: The data comprised hand radiographs of patients undergoing renal dialysis. Normal hand radiographs obtained in trauma patients were assessed as control images. One hundred fifteen images acquired with a photostimulable-phosphor computed radiography system were analyzed. Image selection and initial assessment were by consensus of two experienced radiologists, who graded the radiographic changes of hyperparathyroidism with the Ritz scoring system. The images were then presented to four readers in three formats: hard-copy output and soft-copy presentations at 2K2 and 1K2 resolutions. These readers scored pathologic change and image preference. The results were analyzed with the receiver operating characteristic technique. RESULTS: There was a significant improvement in diagnostic performance for both soft-copy formats relative to the hard-copy format (P < .001). No significant difference in diagnostic performance was found between the two soft-copy formats. There was a significant preference for both soft-copy formats relative to the hard-copy format (P < .01), with the 2K2 soft-copy images preferred to the 1K2 images (P < .01). CONCLUSION: Soft-copy reporting can provide superior diagnostic performance even for images viewed at a modest (1K2) resolution. The lack of difference between the two soft-copy formats has important economic implications with respect to departmental hardware requirements.

Bone and Bones↗

The outcome of intestinal fistulae: the Louisiana State University Medical Center--Shreveport experience.

Fistulae arising from the intestinal tract are associated with significant morbidity and mortality rates. Most contemporary studies of fistulae report mortality rates between 6 and 20 per cent. The major causes of death in these patients are sepsis, electrolyte imbalance, and malnutrition. A total of 48 patients with either external or internal intestinal fistulae were reviewed in this study over a 5-year period at the Louisiana State University Medical Center at Shreveport. Intestinal fistulae were classified into three types, anatomic site, physiologic type, and etiology, to evaluate morbidity and mortality rates. We also attempted to evaluate the role of parenteral nutrition in this patient population, but our data were inconclusive because of the limited number of patients. There was no difference in mortality rates associated with anatomical sites. High-output fistulae were associated with a higher mortality rate compared to low-output fistulae. Fifty-six per cent of the patients achieved closure. The overall mortality rate was 21 per cent. Spontaneous closure rates were lower when compared to those in other studies. This was attributed to sepsis, malignancy, and history of previous radiation therapy. Management of intestinal fistulae includes control of sepsis, correction of electrolyte disturbances, nutritional support, and operative intervention if necessary.

Adolescent↗

Magnetic resonance of the neonatal brain.

We discuss the advantages of magnetic resonance (MR) technique in the study of the neonatal brain. Major results have been obtained concerning the understanding of the normal appearance of the developing brain. Bands of cells migrating to the cortex have been identified in the frontal periventricular white matter up to late gestation. MR can also identify the disappearance of the subependymal germinal matrix with increasing gestational age. With respect to the lesions of prematurity, subependymal germinal matrix hemorrhages were for the first time identified by MR underneath the posterior horns of the lateral ventricles. Subtle ischemic lesions of the periventricular white matter, not detected by brain ultrasound scan, are also described. Analysis of the lesions of the term neonates showed the role of MR in defining a more precise prognosis of infants who have sustained "birth asphyxia". The lesions can affect the basal ganglia, watershed areas of the white matter, and cortex. MR scans performed in the second week of life seem to show a stronger association with the outcome. Brain ischemic areas of the term neonates presenting with focal or multifocal seizures can also be detected by MR. These infarcted zones are usually located in the perfusion territory of the middle cerebral artery, more often in the left hemisphere. The timing of the scan is an important factor as the conventional MR can be negative in the first two-three days after the seizures. The diffusion-weighted imaging (DWI) is a new MR technique very sensitive to acute ischemic injury, and it may solve the problem of the scan timing.

Brain↗