Centrifugal ultrafiltration method for rapid concentration of Legionella pneumophila urinary antigen.
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Antigens, Bacterial↗
Biomedical subjects
J Domínguez
Publications and source records attributed to J Domínguez.
Use of quantitative and semiquantitative procalcitonin measurements to identify children with sepsis and meningitis.
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Bacteremia↗
Expression of porcine CD163 on monocytes/macrophages correlates with permissiveness to African swine fever infection.
Monocytes-macrophages, the target cells of African swine fever virus (ASFV) are highly heterogeneous in phenotype and function. In this study, we have investigated the correlation between the phenotype of specific populations of porcine macrophages and their permissiveness to ASFV infection. Bone marrow cells and fresh blood monocytes were less susceptible to in vitro infection by ASFV than more mature cells, such as alveolar macrophages. FACS analyses of monocytes using a panel of mAbs specific for porcine monocyte/macrophages showed that infected cells had a more mature phenotype, expressing higher levels of several macrophage specific markers and SLA II antigens. Maturation of monocytes led to an increase in the percentage of infected cells, which correlated with an enhanced expression of CD163. Separation of CD163+ and CD163- monocytes demonstrated the specific sensitivity of the CD163+ subset to ASFV infection. In vivo experiments also showed a close correlation between CD163 expression and virus infection. Finally, mAb 2A10 and, in a lower extent, mAb 4E9 were able to inhibit, in a dose-dependent manner, both ASFV infection and viral particle binding to alveolar macrophages. Altogether, these results strongly suggest a role of CD163 in the process of infection of porcine monocytes/macrophages by ASFV.
Animals↗
Immunohistochemical characterisation of PCV2 associate lesions in lymphoid and non-lymphoid tissues of pigs with natural postweaning multisystemic wasting syndrome (PMWS).
The lymphoid, renal, pulmonary, and hepatic lesions of naturally occurring postweaning multisystemic wasting syndrome (PMWS) affected pigs have been studied by means of immunohistology. Ten conventionally reared pigs showing acute clinical signs of PMWS were selected from a farm on which animal were seronegative to porcine reproductive and respiratory virus and to Aujeszky's disease virus. All pigs were positive in tests for porcine circovirus type 2 by ISH and IHC. Monoclonal and polyclonal antibodies to CD3, CD79alpha, CD45RA (3C3/9), lysozyme, SLA-II-DQ (BL2H5), and MAC387 were used to characterise cells in PMWS lesions. The most relevant changes were reduction or loss of B and T lymphocytes, increased numbers of macrophages, and partial loss and redistribution of antigen presenting cells throughout lymphoid tissues compared to uninfected controls. The characteristics of lymphoid lesions in the present study strongly suggest an immunosuppressive effect of PMWS in affected pigs.
Animals↗
Pravastatin decreases cold ischemia--but not alloantigen-dependent transplant arteriosclerosis.
BACKGROUND: Alloantigen mismatch and cold ischemia have been shown to induce transplant arteriosclerosis. Pravastatin (PR) decreases arteriosclerosis probably related to an immunosuppressive effect. Statins possess other nonimmune properties that may be beneficial to transplantation. We studied the effect of PR on cold ischemia and alloantigen-induced transplant arteriosclerosis in syngeneic (SYN) and allogeneic (ALLO) aortic transplantation models. METHODS: Lewis rats served as the donors and recipients for SYN transplants and Brown Norway rats were donors for ALLO transplants. Aortic segments that had been preserved at 4 degrees C in Euro-Collins solution for 0 or 24 hours were transplanted to the infrarenal aorta of the recipients PR (10 mg/kg/d) was administered for 12 weeks prior to morphometric studies. Areas of intimal thickness and its relation to total vessel area were calculated. Lipid levels were measured at 12 weeks. RESULTS: Aorta rings preserved for 24 hours showed marked intimal thickening compared to controls (SYN, CI 0 hours = 21.5% +/- 16.5% vs SYN, CI 24 hours = 50.7 +/- 9.5%, P <.05). PR significantly decreased thickening (SYN, CI 24 hours + PR = 41.7 +/- 12.2 (P <.05) vs SYN, CI 0 hours on SYN, CI 24 hours). There was a nonsignificant decrease in thickening among ALLO transplants treated with PR (ALLO = 31.4 +/- 15.9 vs ALLO + PR = 23.8 +/- 18.8; P >.05). PR had no effect on lipid levels. PR decreases cold ischemia induced transplant arteriosclerosis in this syngeneic aortic transplant model, but does not affect an alloantigen-mediated process. The beneficial effect of PR is not related to its lipid-lowering properties but probably to a nonimmune effect.
Animals↗
A new epitope on swine CD5 molecule detected by monoclonal antibody 5F12/9.
This paper describes the production and characterization of a monoclonal antibody (MAb), 5F12/9, that recognizes a new epitope on porcine CD5. Conformation of its CD5 specificity was obtained by means of sequential immunoprecipitation and Western blot experiments in combination with anti-CD5 MAb 1H6/8, whereas cross-blocking experiments with both MAbs showed that they reacted with different epitopes.
Animals↗
Usefulness of urinary antigen detection by an immunochromatographic test for diagnosis of pneumococcal pneumonia in children.
We evaluated an immunochromatographic assay detecting pneumococcal antigen in urine samples from children diagnosed with pneumococcal pneumonia. The sensitivity and specificity of the immunochromatographic test with nonconcentrated urine (NCU) were 86.7 and 62.9%, respectively; with concentrated urine (CU), they were 100 and 11.7%, respectively. Pneumococcal antigen was also detected in 42.5% of NCU and 87.1% of CU samples from nasopharyngeal carriers. This is a nonspecific test for the diagnosis of pneumococcal pneumonia in children, particularly the very young.
Adolescent↗
Utility of an in-house mycobacteriophage-based assay for rapid detection of rifampin resistance in Mycobacterium tuberculosis clinical isolates.
A rapid in-house mycobacteriophage-based assay to identify multidrug resistance by detecting the rifampin susceptibility of Mycobacterium tuberculosis in a microtiter plate format was evaluated. The sensitivity, specificity, and overall accuracy of the assay were 100%. This test is rapid to perform and suitable for widespread implementation.
Antibiotics, Antitubercular↗
Expression and inheritance of hypersensitive resistance to rice hoja blanca virus mediated by the viral nucleocapsid protein gene in transgenic rice.
Rice hoja blanca virus (RHBV) is a major virus disease of economic importance affecting rice in northern South America, Central America and the Caribbean. This is the first report of transgenic resistance to RHBV and the transformation of an indica rice variety from Latin America. Rice transformed with the RHBV nucleocapsid protein ( N) gene had a significant reduction in disease development. Several reactions were observed that ranged from susceptible to completely resistant plants (immunity). The resistant reactions were characterized by the production of local lesions like a hypersensitive reaction or a recovery phenotype with the emergence of symptom-less new leaves. These transgenic RHBV-resistant rice lines expressed the N gene RNA at low levels that were below the detection limit by Northern blots and only resolved by RT-PCR. The nucleocapsid protein could not be detected in any of the transgenic plants either by Western or ELISA tests. These results suggest that the resistance encoded by the N gene in these plants appears to be mediated by RNA. When challenged with RHBV, the resistant transgenic lines showed a significant increased performance for important agronomic traits including the number of tillers, the number of grains per plant and the yield as compared to the susceptible control. Furthermore, upon inoculation some of the most-resistant transgenic lines showed agronomic traits similar to the uninoculated non-transgenic Cica 8 control. Using both agronomic traits and disease severity as criteria, several of the most-resistant lines were followed through the R(4) generation and demonstrated that the N gene and RHBV resistance was inherited in a stable manner. These transgenic rice lines could become a new genetic resource in developing RHBV-resistant cultivars.
Capsid Proteins↗
Factors related to persistence of Legionella urinary antigen excretion in patients with legionnaires' disease.
The aim of this prospective study was to compare patient characteristics, clinical data, and evolution of Legionella pneumophila pneumonia according to the duration of Legionella urinary antigen excretion. Urine samples from 61 patients with Legionella pneumonia diagnosed by detection of urinary antigen were obtained periodically until urinary antigen could no longer be detected. Cases were divided into two groups based on the duration of urinary antigen excretion: group I (46 patients, <60 days) and group II (15 patients, >or=60 days). Groups were compared for patient characteristics, clinical data, and evolution of pneumonia. Antigen excretion >or=60 days was observed significantly more frequently in immunosuppressed patients ( P=0.001) in whom the time to apyrexia was >72 h ( P=0.002), although only the time to apyrexia remained significant on multivariate analysis ( P=0.006). In conclusion, the duration of Legionella urinary antigen excretion was <60 days in most patients but was longer in immunosuppressed patients with a longer time to defervescence of fever.
Adult↗
Phenotypic characterization of porcine IFN-gamma-producing lymphocytes by flow cytometry.
We have developed a three-colour flow cytometric assay for phenotypic characterization of porcine IFN-gamma-producing lymphocytes. Analyses of activated swine peripheral blood mononuclear cells (PBMC) showed a significant difference in the proportion of IFN-gamma producing cells between young and adult animals (13.2+/-5.8% versus 34.2+/-5.7%). The majority of IFN-gamma producing cells were alphabeta T lymphocytes, although there was also an important proportion of gammadelta T cells particularly in young animals. Within the alphabeta T lymphocytes, the double positive CD4(+)CD8(lo) subset, that contains memory T cells, produced high levels of IFN-gamma, whereas the CD8(hi) T cells ranged from low to high levels of IFN-gamma. Also, consistent with a higher production by memory T cells, the CD45RA(-) subset of both CD4(+) and CD8(+) cells contained higher numbers of IFN-gamma producing cells than the CD45RA(+) subset. Finally, no production of IFN-gamma by either B cells (CD21(+)) or monocytes (SWC3(+)) was detected. This assay may be useful for the assessment of cell-mediated immunity in vaccine trials and may contribute to our understanding of the role of IFN-gamma in protective immunity against important viral diseases of the pig.
Animals↗
[Spontaneous resolution of an acute subdural hematoma].
Acute postraumatic subdural hematoma is known to have bad prognosis. Traditionally, its treatment consisted in urgent surgical evacuation. Rapid spontaneous resolution of the hematoma has been infrequently observed. We report a case of a patient with an acute traumatic subdural hematoma which disappeared without surgery. CT-scan showed a left-sided subdural hematoma with marked mass effect. A low density layer, which implied involvement of CSF in the subdural space, was observed between the hematoma and the inner wall of the skull. We think that the spontaneous resolution of the hematoma was due to the dilution and washing-out by the CSF through an arachnoid tear. Another suggested mechanism in the literature is blood redistribution in the subdural space. Selected patients with an acute subdural hematoma can be managed with conservative treatment.
Aged↗
Steroidal management and serum cytokine profile of a case of alcoholic hepatitis with leukemoid reaction.
Leukemoid reactions (LRs) are rare in alcoholic hepatitis (AH), but they are a sign of poor prognosis. The treatment of AH with corticosteroids is controversial, though several reports suggest that these should be used in severe cases of AH. We report a case of AH-associated LRs that presented with an increase of the serum concentrations of the proinflammatory cytokines interleukin (IL)-18 (an initiator of inflammation) and IL-1beta (likely responsible for the neutrophilia of the LRs). These findings provided a pathogenic indication for the use of corticosteroids (that block the transcription of IL-1beta), and this approach achieved a clinical and analytical recovery in our patient. This pathogenic mechanism might also underlie other cases of LRs and other complications of AH, thus providing a rationale for the benefits of corticotherapy in these rare but severe conditions.
Adult↗
[Intracranial hypertension associated with type I Chiari malformation].
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Adult↗
Changes in peripheral blood leukocyte populations in pigs with natural postweaning multisystemic wasting syndrome (PMWS).
The objective of the present study was to analyze, by flow cytometry, changes in PBMC subsets in pigs having postweaning multisystemic wasting syndrome (PMWS), a new condition associated to porcine circovirus type 2 (PCV2) infection. Thirteen acutely PMWS affected pigs were selected from a farm seronegative to porcine reproductive and respiratory syndrome virus (PRRSV) and to Aujeszky's disease virus (ADV); 11 clinically healthy pigs were selected from a high health farm with no history of PMWS and free of the major swine pathogens, and used as a control group. All pigs were necropsied, and tissue samples were fixed in formalin; blood with EDTA anticoagulant was used to perform the flow cytometric analysis. PBMC were incubated with mAb against porcine CD3, CD4, CD8, CD25, CD45, IgM, SWC3, and SLA-Class II. Flow cytometric analysis showed substantial changes in leukocyte subsets in the peripheral blood of PMWS-affected pigs, which were characterized by an increase of monocytes, a reduction of T (mainly CD4(+)) and B-lymphocytes, and the presence of low-density immature granulocytes. Altogether, these changes would suggest an inability of acutely PMWS-affected pigs to mount an effective immune response.
Animals↗
Workshop studies on monoclonal antibodies in the myeloid panel with CD11 specificity.
Several putative anti-human and swine CD11-specific monoclonal antibodies (mAbs) were included in the myeloid section of the Third International Swine CD Workshop. Failure of clustering analysis to group these mAbs together prompted additional analyses to define the specificities of these mAb. Combination of one and two-color flow cytometry (FCM) and immunoprecipitation (IP) allowed the definition of the mAb into three CD11 groups. Cellular distribution of the molecules recognized by anti-human CD11b and c mAbs on swine cells proved to be significantly different from that found in humans.
Animals↗
Rapid detection of several mycobacterial species using a polymerase chain reaction reverse hybridisation assay.
The ability of a polymerase-chain-reaction-based technique combined with a reverse hybridisation line-probe assay to detect and identify eight of the most clinically significant mycobacterial species directly from cultures in liquid medium was evaluated. The line-probe assay allows simultaneous identification of eight different mycobacterial species. Ninety-seven mycobacterial strains belonging to 22 different species were evaluated. All strains were previously inoculated into MB/BacT bottles (Organon Teknika, USA). The sensitivity and specificity of the test when applied on positive MB/BacT liquid cultures containing isolates from mycobacterial species included as specific probes on the line-probe assay strip was 100% and 100%, respectively. These results further support the potential clinical usefulness of this technique in the diagnosis of mycobacterial infections.
Bacterial Typing Techniques↗
Immunohistological study of the immune system cells in paraffin-embedded tissues of conventional pigs.
The distribution of different cells of the immune system has been studied in formalin-fixed paraffin-embedded tissues from conventionally reared healthy pigs, using immunohistological techniques. The samples collected were: lungs, tonsils, lymph nodes (mediastinal, mesenteric, inguinal and submandibular), pancreas, spleen, liver, kidney, adrenal gland, ileum and stomach. A total of six primary antibodies anti-CD3, anti-CD79alpha, Mac 387, anti-lysozyme, anti-CD45RA (3C3/9) and anti-SLA-II-DQ (BL2H5) were used with a standard avidin-biotin peroxidase (ABC) method. Anti-CD3 and anti-CD79alpha mAb-reacted, respectively with cells located in T cell areas and B cell areas. Mac 387 recognised circulating polymorphonuclear leukocytes, while anti-lysozyme-stained resident macrophages in all tissues. 3C3/9 and BL2H5, were assessed in formalin-fixed paraffin-embedded tissues for the first time. 3C3/9 identified B lymphocytes, in primary follicles and mantle zones, a subpopulation of T cells, especially located in the marginal zone of the spleen and a variable number of immunoblasts, in the germinal centres. BL2H5 reacted with B cells in the mantle zones of the follicles of lymphoid tissues, with dendritic and interdigitating cells in all studied lymphoid tissues and with a variable number of resting and activated T cells in the periarteriolar lymphoid sheath (PALs), marginal zone and red pulp of the spleen. Furthermore, it stained Kupffer and perivascular macrophages in the liver. This study represents a detailed histological study of the distribution of the most important subpopulations of immune system cells in conventional, healthy pigs. In our view, these tools should be useful for future comparative studies in disease conditions.
Animals↗