Subsistence swine farming: seroprevalence and risk factors associated with Lawsonia intracellularis infection in the state of Minas Gerais Brazil in 2016.

Although it is considered an economically relevant and prevalent disease, little information is available on the epidemiology and risk factors of porcine proliferative enteropathy (PPE) in commercial pigs, and no publication is available on subsistence pig farming. The objectives of this study were to estimate the seroprevalence of L. intracellularis and identify associated risk factors in backyard pigs in the 12 mesoregions of the state of Minas Gerais, Brazil. Blood from pigs between 2 months and 6 years of age were sampled; an epidemiological questionnaire was applied to 288 properties investigated in 2016. Serum samples were tested for the presence of anti-L. intracellularis antibodies using an immunoperoxidase monolayer assay. The seroprevalence of L. intracellularis was 97.7% (CI 95%: 96.7-98.4), and there was no statistical difference among the prevalence of the sampled mesoregions. Only 3 of the 12 risk factors were significant when samples were analyzed from strongly seropositive animals (≥ 1:120) in a Poisson multivariate regression model. There was an interaction between properties in peri-urban areas and extensive production systems. This interaction demonstrated an increase in prevalence rates by 3.7 times (95%CI: 2.4-5.8). Properties close to dumps demonstrated an increase in prevalence rates by 2.2 times (95%CI: 0.99-4.8). In conclusion, anti-L. intracellularis antibodies were widely dispersed in subsistence pig farming's in Minas Gerais, indicating a wide circulation of the agent in this type of production system. The interactions of animals raised close to peri-urban areas, extensively, and close to landfills are risk factors for spread of PPE.

Clinical consequences of using PNA-FISH in Staphylococcal bacteraemia.

To optimize patient treatment and rational use of antimicrobials, it is important to provide fast information on findings in blood-cultures (BCs). The purpose of this study was to evaluate the impact of using peptide nucleic acid fluorescence in situ hybridization (PNA-FISH) on positive BCs containing Gram-positive cocci in clusters to differentiate between Staphylococcus aureus (SA) and coagulase negative staphylococci (CoNS) on the prescribed antimicrobial therapy and on the number of contacts between microbiologist and clinician. All cases of positive BCs in our laboratory with SA or CoNS in the year 2011 were identified and the charts were reviewed retrospectively. The group of patients with BCs tested with PNA-FISH was compared to the group of patients with untested BCs. A total of 200 patients with SA and 725 patients with CoNS were included. The mean number of contacts was 0.82 when PNA-FISH showed CoNS and 1.39 when PNA-FISH was not done (p < 0.0001). More patients were recommended appropriate antimicrobial therapy for SA bacteraemia in the PNA-FISH group (98.0%) than in the non-PNA-FISH group (89.4 %) (p = 0.025). The percentage treated with dicloxacillin was 29.6 in the PNA-FISH group, and 8.2 in the non-PNA-FISH group (p = 0.0003). The use of PNA-FISH on BCs in this study was associated with more appropriate and narrow spectrum antimicrobial therapy for patients with SA in an area with low prevalence of methicillin-resistant SA, and a lower number of contacts between clinical microbiologist and clinician about BCs with CoNS as contaminants.

Specific protein content of pools of plasma for fractionation from different sources: impact of frequency of donations.

BACKGROUND AND OBJECTIVES: Plasma pools for the production of human plasma medicinal products are distinguished according to the collection method (recovered or apheresis plasma) and the donor remuneration status. National regulations and the physical status of the donor determine the donation frequency and plasma volume per session. Relevant protein contents of different types of pools have not fully been compared. MATERIALS AND METHODS: We compared the levels of total protein, 15 main relevant plasma protein markers, and anti-B19 and anti-Streptococcus pneumoniae IgG in single-type pools of donations from different countries (Belgium, Finland, France, the Netherlands, Germany, United States). Both recovered plasma from non-remunerated donors and apheresis plasma from remunerated and non-remunerated donors were studied. RESULTS: Pools from paid US high-frequency, high-volume plasmapheresis donors showed significantly lower total protein (-9%), albumin (-15%), total IgG (-24%), IgM (-28%), hemopexin (-11%) and retinol-binding protein (-10%) but higher C1-inhibitor, pre-albumin and C-reactive protein contents than pools from unpaid European Union (EU) or US whole-blood or plasmapheresis donors. In contrast to pools from compensated EU plasmapheresis donors, pools from unpaid whole-blood or plasmapheresis donors showed no significant differences, whatever the collection method or country. Reductions in specific protein contents correlated well with protein half-life. CONCLUSION: These results should be taken into account with regard to donor health management and protein recovery.

Degradation of collagen and proteoglycan by macrophages and fibroblasts. Individual potentialities of each cell type and cooperative effects through the activation of fibroblasts by macrophages.

Fibroblasts and macrophages of various sources (peritoneal, alveolar or bone marrow-derived), from either rabbit or mouse, were cultured, independently or together, at the surface of [3H]proteoglycan/[14C]collagen-coated plates to evaluate their capacities for proteoglycan and collagen degradation. The various macrophage populations differed widely in their potentialities for proteoglycan and particularly, for collagen degradation, native collagen being significantly degraded, in this model only by rabbit alveolar macrophages. Fibroblasts were as active in proteoglycan degradation as the most active macrophage preparations, but their potential for collagen degradation appeared much higher than that of macrophages. Moreover, all types of macrophages secreted a factor, a monokine, that activated collagen and proteoglycan degradation by fibroblasts. Thus, fibroblasts might well be a major effector cell, active in connective tissue degradations occurring under chronic inflammatory situations.

A multiple transgenic mouse model with a partially humanized activation pathway for helper T cell responses.

Mice expressing human CD4 and human MHC II molecules provide a valuable model both for the investigation of the immunopathogenetic role of human autoantigens and for the development of therapeutic strategies based on modulating helper T cell activation in vivo. Here we present a novel mouse model expressing HLA-DR17 (a split antigen of HLA-DR3) together with human CD4 in the absence of murine cd4 (CD4/DR3 mice). Human CD4 accurately replaces murine cd4 within T cells. In particular, the preservation of cd8(+) and CD4(+) T cell subsets distinguishes CD4/DR3 mice from other multiple transgenic models in which the alternative T cell subsets are fundamentally disturbed. Moreover, human CD4 is also faithfully expressed on antigen presenting cells such as dendritic cells and monocyte/macrophages, so that the overall transgenic CD4 expression pattern resembles very closely that of humans. HLA-DR3 expression in the thymus correlates very closely to that of mouse MHC II. In contrast, only 70% of mouse MHC II positive cells in spleen, lymph node, and peripheral blood coexpress HLA-DR3. No significant bias was found with regard to particular leucocytes in this respect. The stimulation of helper T cells clearly depends on the interaction between the human transgene products, since mAbs to HLA-DR and/or CD4 completely blocked in vitro recall responses to tetanus toxoid. CD4/DR3 mice represent a partially humanized animal model which will facilitate studies of DR3-associated autoimmune responses and the in vivo determination of the therapeutic potential of mAbs to human CD4.

Modification of factor VIII in therapeutic concentrates after virus inactivation by solvent-detergent and pasteurisation.

The addition of a pasteurisation step to a solvent/detergent (SD) treated FVIII concentrate has recently resulted in enhanced inhibitor incidence in patients in Germany and Belgium. We have investigated the effect of virus inactivation procedures on FVIII function by preparing experimental concentrates from the same starting cryoprecipitate with the following procedures: none (N); dry heat (DH); pasteurisation (P); solvent/detergent (SD); solvent detergent + dry heat (SDDH); solvent detergent + pasteurisation (SDP). In addition, several clinical SD concentrates with and without pasteurisation were studied. There were no significant differences in fibrinogen and vWF content and in the ratio of one-stage/chromogenic FVIII activity among any of the samples studied. In thrombin proteolysis and FXa generation experiments, there were no differences in results on samples N, DH, P, and SDDH from those on sample SD. However sample SDP gave markedly different results from sample SD in the following respects: slower thrombin proteolysis (t(1/2) = 12.0 min vs 1.9 min); more rapid FXa generation (rate 2.5 times that of SD); enhanced phospholipid binding (K(D) = 3.89 x 10(-11) M vs 5.53 x 10(-10) M). Similar differences between SDP and SD were seen in the clinical samples. The observed changes in the FVIII activity occurred in combination with SD and pasteurisation, but not with either treatment alone. These results suggest that SDP treatment may enhance exposure of the phospholipid binding site in the C2 domain of FVIII, and since inhibitors to the SDP product are predominantly against C2, these findings could be relevant to the enhanced immunogenicity of the SDP product.

Factor VIII inhibitors in previously treated haemophilia A patients with a double virus-inactivated plasma derived factor VIII concentrate.

Antibodies to factor VIII (inhibitors) are usually produced at the beginning of treatment with factor VIII and are rare in multitransfused patients. Such antibodies are deemed to be patient-related, as supported by the description of a number of associated risk factors. However, a second category of inhibitors has recently been identified, namely antibodies occurring in multitransfused patients as a result of exposure to a particular factor VIII concentrate. A first outbreak of product-related inhibitors was recently described. The present paper describes the second well-documented occurrence of such inhibitors. Eight out of 140 multitransfused patients with severe haemophilia A developed an inhibitor to factor VIII shortly after changing treatment to a double-virus inactivated plasma-derived factor VIII concentrate. In addition to solvent-detergent treatment, this concentrate was pasteurised at 63 degrees C for 10 hours. Exposure to the pasteurised product before inhibitor detection ranged from 9 to 45 days. Inhibitor titers varied between 2.2 and 60 Bethesda Units and recovery of transfused factor VIII ranged from 0.21 to 0.68 (expressed as i.u./dl factor VIII rise per i.u./kg administered). In contrast to usual inhibitors in haemophilia A patients, these product-related inhibitors showed complex inhibition kinetics. They were found specific for the factor VIII light chain. The inhibitors gradually declined when exposure to the pasteurised product was stopped, despite further treatment with other factor VIII concentrates. The present data stress the importance of carefully monitored clinical studies, both in previously treated and previously untreated patients, before introduction of a new or modified clotting factor concentrate.

Inhibitors in German hemophilia A patients treated with a double virus inactivated factor VIII concentrate bind to the C2 domain of FVIII light chain.

To reduce the risk of transmission of hepatitis A virus, an Octapharma produced factor VIII (fVIII) concentrate treated with solvent detergent (FVIII-SD) was further pasteurized after purification. This product, Octavi SDPlus (FVIII-SDP), was marketed in Europe in 1993 to 1995. Inhibitors appeared from September to October, 1995, in 12 of 109 previously treated German hemophilia A patients. A study of similarly treated Belgian patients, who also developed inhibitors, had shown antibodies to the fVIII light chain (domains A3-C1-C2) only. In the present study, the epitope specificity of 8 German inhibitor plasmas was also found to be restricted to the light chain. In radioimmunoprecipitation assays to localize the light chain epitope(s), antibody binding to heavy chain (domains A1-A2-B) was 11-148 fold lower than to the C2 domain, and binding to recombinant A3-C1 was barely detectable. These results were supported by >95% neutralization of a high responder inhibitor titer by the C2 domain.

Cellular pharmacology of deferrioxamine B and derivatives in cultured rat hepatocytes in relation to iron mobilization.

Two radiolabelled derivatives of deferrioxamine B (DF) have been synthesized: methyl-DF and acetyl-DF. Both derivatives are non cytotoxic and stable in cell culture but they are degraded in human plasma and more extensively in rat plasma. Methyl-DF, acetyl-DF and DF mobilize radioiron to the same extent from hepatocytes loaded with 59Fe citrate in the same range of extracellular concentrations. The uptake and release of the 3H-labelled derivatives and their corresponding iron complexes have been measured and appear to represent a passive phenomenon resulting from the gradient of concentration between the cellular compartment and the extracellular medium. The results indicate that only a limited pool of cellular iron is accessible for chelation and that neither the permeability of the cellular membrane, nor the intracellular concentration of the chelators are the limiting factors for iron mobilization. On the basis of the subcellular distribution of the 3H-DF analogues, methylamine inhibition of iron chelation by siderophores in cell cultures and the positive effect of acidic pH and hydrolysis by lysosomal enzymes on in vitro iron mobilization from radiolabelled ferritin, we suggest that iron mobilization by DF and its derivatives occurs in lysosomes where they complex iron released from ferritin under the conjugate actions of acidic pH and lysosomal enzymes.

Main Drain virus as a cause of equine encephalomyelitis.

Main Drain virus, which is thought to be transmitted normally among rabbits and various rodents by its natural vector, Culicoides variipennis, was isolated repeatedly from brain tissue of a sick horse from Sacramento County, California, and was implicated as the causative agent. Signs of illness were incoordination and ataxia, stiff neck, head pressing, inability to swallow, fever, and tachycardia.

Visualizing inflammation activity in rheumatoid arthritis with Tc-99 m anti-CD4-mAb fragment scintigraphy.

PURPOSE: T-cell-located CD4 antigen represents one of the therapeutic targets in rheumatoid arthritis (RA). However, up to now there is no established imaging tool to visualize this target in vivo. The aim of our study was to assess the safety and tolerability of a technetium-99m labelled murine anti-human CD4 IgG1-Fab fragment ([(99m)Tc]-anti-CD4-Fab, [(99m)Tc]-EP1645) in patients with active synovitis due to RA, and to evaluate its potential as a marker of disease activity. METHODS: In the present phase I proof of principle study five patients with RA were examined. Planar scans of the whole body, hands, and feet were taken 30 min up to 24h after application of 550 ± 150 MBq [(99m)Tc]-anti-CD4-Fab, followed by visual analyses, comparison with clinical data in 68 joints per patient and semiquantitative analysis of hand and wrist joints. RESULTS: Neither infusion related adverse events nor adverse events during follow up were observed. No increase in human anti-murine antibody titres was seen. All patients had positive scans in almost 70% of clinically affected joints. Positive scans were also found in 8% of joints without evidence of swelling or tenderness. CONCLUSION: Scintigraphy with [(99m)Tc]-anti-CD4-Fab is a promising technique for evaluation of inflammatory activity in patients with RA, pre-therapeutical evaluation of CD4 status and therapy control. Tracer uptake in clinically inconspicuous joints strongly indicates diagnostic potential of [(99m)Tc]-anti-CD4-Fab. Whether this technique is eligible as a prognostic factor in RA needs to be analysed in further studies as well as the pathophysiological background of clinically affected joints lacking tracer uptake.

[Mechanism of action and therapeutic use of cyclosporin A. Experimental and clinical experiences in dermatology]. / Wirkungsweise und therapeutische Anwendung von Cyclosporin A. Experimentelle und klinische Erfahrungen in der Dermatologie.

The clinical application of cyclosporine A caused many success in transplantation surgery as well as in the treatment of (auto-) immune disorders. The biological highly active substance differs from conventional immunosuppressive drugs (i.e. steroids, azathioprine) by it's selective action on T-lymphocytes. However, current experimental findings indicate a direct influence on epidermal keratinocytes, too. First therapeutical experiences, concerning systemic and topical application in different skin diseases are summarized.

[Practical experiences in the application of the BL-series of monoclonal antibodies for phenotyping of lymphocytes]. / Praktische Erfahrungen zum Einsatz monoklonaler Antikörper der BL-Serie für Lymphozytenphänotypisierungen.

Determination of peripheral lymphocytes has increasing importance in laboratory examinations. However, evidence concerning differential diagnosis is low. For experimental questions and therapeutical monitoring application of this method is more distributed. This paper transfers practical experiences concerning the detection of cellular surface antigens. For the investigations, monoclonal antibodies from the Karl-Marx-University Leipzig were used in immunofluorescence microscopy. Limits of the test are demonstrated (in skin diseases and HIV-infection f.e.).

[Anti-epidermal cytotoxic reactions in the effector stage of lichen ruber planus]. / Antiepidermale zytotoxische Reaktionen in der Effektorphase des Lichen ruber planus.

Immunohistochemical investigations on skin lesions of lichen ruber planus indicate HLA-DR-expression by keratinocytes, Langerhans cells (LC), and infiltrated mononuclear cells. In the early stage of this condition, the lymphocytic infiltrate consists of a considerable part of (CD8+) cytotoxic/suppressor- and (LGL+) natural killer cells. Evidently, the cytotoxic potential of these lymphocyte subpopulations is directed against basal keratinocytes and epidermal LC. As a result of cytotoxic/cytolytic activities, non-cell associated CD 1-reactivity accumulates in the dermoepidermal junction zone, thus indicating damaged LC.

[Changes in the functional surface markers of lymphocytes in short-term culture]. / Veränderungen funktioneller Oberflächenmerkmale von Lymphozyten in der Kurzzeitkultur.

Cultured human lymphocytes are a suitable system for testing pharmacological, allergological, and toxicological effects of substances in vitro. For such investigations, the knowledge about the behaviour of the test system under standardized conditions is an important prerequisite. In the 72 h short-time-culture, no changes in T-lymphocytes were found. Small decreases occurred in the subpopulations of cytotoxical/suppressor cells, and inducer/helper cells, respectively. Number of cells expressing class II antigens of the main histocompatibility complex decreases, too (to 62%). Tritium thymidine incorporation rates inform about DNA-synthesis.

[Demonstration of epidermal Langerhans cells]. / Zur Darstellung epidermaler Langerhanszellen.

Langerhans Cells (LC) are centrally involved in the induction of skin-located immunological processes, including kontactallergical reactions. Because of the growing interest in LC functions, and dynamics, various possibilities for their detection reaching from histological staining up to immunohistochemical characterization by means of monoclonal antibodies, have been developed. The concern of this paper is to overview different visualization techniques.

Immunological relatedness of ribosomes from mycobacteria, nocardiae and corynebacteria, and microorganisms in leprosy lesions.

Serological relatedness of ribosomes from microorganisms of the Mycobacterium, Nocardia, and Corynebacterium genera has been analyzed by the microplate immunodiffusion technique. Mycobacterium and Nocardia proved homogeneous and closely related taxa, whereas Corynebacterium was found to be a heterogeneous phylum connected by remote links to the others. The taxonomic position of "diphtheroid microorganisms" (non-acid-fast, gram-positive bacteria morphologically similar to corynebactria), which were found together with Mycobacterium leprae in human leprosy lesions, was also investigated. Ribosomes of diphtheroid bacteria strongly cross-reacted with antisera against several mycobacteria and nocardiae but not against corynebacteria. Moreover, ribosomes from independently isolated diphtheroid strains proved serologically related and yielded strong cross-reactions with antisera against M. leprae as well as with sera from leprosy patients. Hence, diphtheroid microorganisms represent a homogeneous group immunologically related to mycobacteria in general and more specifically to M. leprae.