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No description available.

No description available.

No description available.

Infection of IFNγ receptor knockout (IFNγ R-/-) mice with murine gammaherpesvirus-68 (MHV-68) results in fibrosis in the lung, spleen, liver and lymph nodes. In the spleen, pathology involves an increase in the number of latently infected B cells that corresponds with a Th2 biased immune response, in which germinal centres become walled off and fibrosis dominates the splenic architecture. Remarkably, the spleen recovers from this pathology, and the starting point for this process is a loss of latently infected B cells. The aim of this project is to gain further understanding of the control of MHV-68 latent infection in the absence of IFNγ response. This project investigates: (1) the mechanisms that result in the loss of splenocytes, in particular the reduction of latently infected B cells; (2) the dynamics of macrophages in the induction, expression and recovery of fibrosis. Several approaches were employed to examine the hypothesis that the massive cell loss in IFNγR-/- spleen is caused by apoptosis. However, there was no evidence for excessive apoptosis throughout the development of fibrosis. Moreover, RT-PCR analysis showed that there was no significant increase in expression of viral genes associated with lytic infection. Hence it is unlikely that viral reactivation and subsequent lytic infection occurs. These data suggest apoptosis and viral reactivation are not the main mechanisms that cause splenic cell loss. Furthermore, B cell subpopulations and cells that express viral ORF73 in IFNγR-/- mice were examined using a recombinant virus. The ORF73-expressing cells are mainly germinal centre B cells and memory B cells. These two subpopulations undergo a drastic decrease in numbers during fibrosis, whereas naïve B cells, which are less susceptible to infection, maintain a relatively stable population. Therefore, the significant reduction of latently infected B cells appears to be related to the removal of germinal center B cells and memory B cells. Macrophages induced by Th2 cytokines are considered to be pro-fibrotic, and they are reported to have the potential to differentiate into myofibroblasts. In order to determine the role played by macrophages in MHV-68 induced fibrosis, transgenic mice with eGFP constitutively expressed in macrophages and dendritic cells were used. A different pattern of macrophage distribution in IFNγR-/- mice was observed compared to that in wild type mice. Moreover, the number of splenic macrophages changed dramatically in the spleen at different stages of fibrosis. The possibility that alternatively activated macrophages differentiate into myofibroblasts was investigated by co-staining with α-SMA antibody. However, no evidence was found that macrophages are one of the origins of myofibroblasts. This suggests macrophages may play other roles in regulating fibrosis rather than contributing directly to the formation of fibrosis.

No description available.

(1) Human trypsin was purified from human pancreatic tissue and used as an immunogen in the production of monoclonal antibodies. It was also used as a standard preparation of human trypsin in the screening of monoclonal antibodies and in the development of an immunoradiometric assay (IRMA). (2) Eight monoclonal antibodies were raised to human trypsin, two of which were subsequently employed in the development of an IRMA to measure immunoreactive trypsin (IRT) in blood and blood spots. (3) An antiserum to human trypsin was raised in sheep and employed in a sandwich ELISA. (4) Monoclonal antibodies were tested by a number of different screening systems including ELISA, immunoblotting and immunoadsorption assays with 125I-labelled human trypsin. (5) Six selected monoclonal antibodies were tested for use (a) as the solid phase antibody and (b) as the radioactive tracer in an IRMA, and against each other for compatability in an IRMA. (6) One combination of antibodies was chosen and experiments performed to determine (a) lack of competition of binding to antigen between the two and (b) positive binding of the combination to serum IRT. (7) An IRMA to measure IRT in blood and blood spots was developed using two monoclonal antibodies, 56/C5/33 (IgG2a) and 125I-labelled 55/A4/31 (IgM). (8) The detection limit of the assay with purified cationic human trypsin as the analyte was determined to be 0.8 ng trypsin. (9) The IRMA was performed with reconstituted blood containing standards of human trypsin (either blood or dried blood spots on filter paper). (10) There is potential for this assay to be further developed and employed as a routine screening assay to detect elevated concentrations of IRT in dried blood spots from newborn infants with cystic fibrosis.

The S100 zinc-binding protein, calprotectin, is a noncovalently associated anionic complex which incorporates two immunologically distinct polypeptides in a molecule of Mr 36.5 kDa. Calprotectin's abundance in neutrophils, where it constitutes 60% of cytosolic protein, suggests an innate host defence function. In vitroit is microbiostatic. The trace metals, iron and zinc, are essential to the growth of most microorganisms. During the acute phase response to infection in man, the marked decrease in plasma iron is associated with a reduction in microbial growth. The similar, though smaller decrease in plasma zinc during this response to infection, may also act as a protective measure. As most microorganisms appear not to have zinc retrieval agents analogous to siderophores for iron retrieval, pathogenic microorganisms may be more susceptible to zinc deprivation than to iron deprivation. Thus, we hypothesized that calprotectin's in vitro candidastatic activity may be mediated by zinc chelation and that this phenomenon may also occur, in vivo, in plasma. In Sabouraud's culture medium, we demonstrated that calprotectin, at concentrations as low as 10ml, has potent, pH dependent candidastatic activity associated with zinc chelation. In plasma from children with cystic fibrosis (CF) & infected children, we demonstrated increased plasma calprotectin concentrations (median: 1832 & 4753 g/l respectively) compared to controls (median: 685 g/l). Plasma calprotectin concentration correlated positively with serum C- reactive protein in infected children, reflecting an association with the acute phase response. In CF children, there were positive correlations between plasma calprotectin and plasma copper and between plasma calprotectin and the Northern score, reflecting associations with the acute phase response and the extent of lung involvement respectively.

La tuberculosis (TB) es una enfermedad social por excelencia, multifactorial y está vinculada estrechamente a la pobreza, hacinamiento, tugurización, desnutrición, al hambre y demás determinantes sociales en nuestro país. La radiografía de tórax es suficiente en muchos casos para su detección, sin embargo aproximadamente en un 21% de los afectados estas sean normales, por ello se sugiriere que la Tomografía computada debe practicarse en los pacientes con sospecha de Tuberculosis pulmonar y radiografía normal o no concluyente. Para esto se incluyeron a 90 pacientes con diagnóstico de TBC confirmada por estudio bacteriológico, los cuales se realizaron una tomografía en el Hospital Nacional Hipólito Unanue entre los meses de julio y diciembre del 2014. En los cuales se evaluó las manifestaciones tomográficas en relación a Tuberculosis pulmonar activa e inactiva. Los hallazgos más frecuente de una tuberculosis pulmonar activa son la zona de consolidación, el patrón micronodular, las adenopatías mediastinales así como el patrón en vidrio esmerilado, con altas frecuencias. Además, vale mencionar la asociación que existe entre los distintos patrones de actividad con el derrame pleural. En caso de la tuberculosis inactiva es muy frecuentemente observar la fibrosis pulmonar asociada a adenopatías calcificadas, bronquiectasias, entre los más representativos. Por lo que se concluye que la Tomografía convencional de pulmones resulta ser una técnica de alta sensibilidad, superior a la radiografía para la detección de casos de tuberculosis activa como inactiva. Palabras clave. Tuberculosis pulmonar. TBC activa. TBC inactiva. Tomografía computada. Zona de consolidacion. Fibrosis. / --- The tuberculosis (TB) is a social excellent disease, multifactorial and is linked narrowly to the poverty, accumulation, tugurización, malnutrition, to the hunger and other social determinants in our country. The radiography of thorax is sufficient in many cases for his detection, nevertheless approximately, in 21 % of the affected ones, these are normal, by it will be suggested that the calculated Tomography must practice him in the patients with suspicion of pulmonary Tuberculosis and normal or not conclusive radiography. For this there were included to 90 patients by TBC's diagnosis confirmed by bacteriological study, which carried out a tomography in the National Hospital Hipólito Unanue between July and December 2014. In which the manifestations were evaluated tomografics in relation to pulmonary active and inactive Tuberculosis. The findings more frequent of a pulmonary active tuberculosis are the zone of consolidation, the micronodular boss, the mediastinal adenopatías as well as the boss in burnished glass, with high frequencies. In addition, it is worth mentioning the association that exists between the different bosses of activity with the pleural spillage. In case of the inactive Tuberculosis, it is very frequently to observe the pulmonary fibrosis associated to calcified adenopatías, bronquiectasias, between the most representative. For what concludes that the conventional Tomography of lungs turns out to be a technology of high sensibility, superior to the radiography for the detection of cases of active tuberculosis or inactive. Keywords: Pulmonary tuberculosis. Active TB. Inactive TB. Computed tomography. Consolidation zone. Fibrosis.

MSc (Med), Faculty of Health Sciences, University of the Witwatersrand / Introduction: Systemic sclerosis (SSc) is a complex autoimmune disease characterised by autoantibody release, leading to microvascular injury, fibroblast activation and increased production of collagen. The genetics of SSc is complex with many genes implicated in the development and maintenance of the extracellular matrix (ECM). The main aim of this study was to test for differential expression of matrix metalloproteinase 1 (MMP1), tissue inhibitor of metalloproteinase 1 (TIMP1) and hepatocyte growth factor (HGF) in SSc patients compared to healthy control individuals and to assess whether the differential expression of these genes could have an impact on clinical features of the disease. Methods: Two skin biopsies were analysed for each of 16 black SSc patients, one from clinically involved skin (lateral forearm) and one from clinically uninvolved skin (back). One skin sample was obtained from 15 ethnically matched control individuals. The differential expression of MMP1, TIMP1 and HGF in the clinically involved and uninvolved patient samples would be compared to control individuals using relative quantification polymerase chain reaction (qPCR). The gene expression profiles were then compared to specific clinical features to deduce whether any of the gene expression profiles is correlated with the manifestation of specific clinical features. Results: MMP1 gene expression was significantly decreased in SSc patients for both involved (p=0.0004) and uninvolved skin (p=0.0004) compared to controls. Conversely, TIMP1 gene expression was significantly increased in SSc patients at both sites compared to controls (p=<0.00001 for both comparisons). A trend of significance was observed for the difference in TIMP1 expression between the involved an uninvolved skin within the patients (p=0.05) with a greater increase in involved skin. HGF had increased gene expression in the patients compared to controls for involved and uninvolved skin (p=0.002 and 0.004, respectively). The difference in gene expression between the involved and uninvolved biopsies was not significant for either MMP1 or HGF (p=0.87 and 0.83, respectively). The only correlates that may have a biological significance are HGF in involved skin correlated with disease activity (r=0.60; p=0.013) and HGF in uninvolved skin vi correlated with skin score (MRSS) with r=0.50 and p=0.048. With regards to the categorical data, two marginally significant observations were found, once again with HGF, which was found to be associated with gender in involved skin (p=0.037) and renal disease in uninvolved skin (0.031). Conclusion: The relative under expression of MMP1 and over expression of TIMP1 reflect the pro-fibrotic state of scleroderma skin. The over expression of HGF suggests that HGF may play a compensatory anti-fibrotic role, although this is not sufficient to overcome the pro-fibrotic state of the skin. This study provides supporting evidence to debunk the myth of uninvolved skin in SSc patients. The altered expression of MMP1, TIMP1 and HGF in the clinically uninvolved skin of SSc patients suggests that all subcutaneous tissue is affected, although to a greater extent in the clinically involved skin of the patients.