Association of Bartonella spp bacteremia with Chagas cardiomyopathy, endocarditis and arrythmias in patients from South America

Infection with Bartonella spp may cause cardiac arrhythmias, myocarditis and endocarditis in humans. The aim of the present study was to evaluate a possible association between Bartonella spp bacteremia and endocarditis, arrhythmia and Chagas cardiomyopathy in patients from Brazil and Argentina. We screened for the presence of bacterial 16S rRNA in human blood by PCR using oligonucleotides to amplify a 185-bp bacterial DNA fragment. Blood samples were taken from four groups of subjects in Brazil and Argentina: i) control patients without clinical disease, ii) patients with negative blood-culture endocarditis, iii) patients with arrhythmias, and iv) patients with chronic Chagas cardiomyopathy. PCR products were analyzed on 1.5% agarose gel to visualize the 185-bp fragment and then sequenced to confirm the identity of DNA. Sixty of 148 patients (40.5%) with cardiac disease and 1 of 56 subjects (1.8%) from the control group presented positive PCR amplification for Bartonella spp, suggesting a positive association of the bacteria with these diseases. Separate analysis of the four groups showed that the risk of a Brazilian patient with endocarditis being infected with Bartonella was 22 times higher than in the controls. In arrhythmic patients, the prevalence of infection was 45 times higher when compared to the same controls and 40 times higher for patients with Chagas cardiomyopathy. To the best of our knowledge this is the first report of the association between Bartonella spp bacteremia and Chagas disease. The present data may be useful for epidemiological and prevention studies in Brazil and Argentina.64465

Identification of metalloprotease gene families in sugarcane

Metalloproteases play a key role in many physiological processes in mammals such as cell migration, tissue remodeling and processing of growth factors. They have also been identified as important factors in the patho-physiology of a number of human diseases, including cancer and hypertension. Many bacterial pathogens rely on proteases in order to infect the host. Several classes of metalloproteases have been described in humans, bacteria, snake venoms and insects. However, the presence and characterization of plant metalloproteases have rarely been described in the literature. In our research, we searched the sugarcane expressed sequence tag (SUCEST) DNA library in order to identify, by homology with sequences deposited in other databases, metalloprotease gene families expressed under different conditions. Protein sequences from Arabidopsis thaliana and Glycine max were used to search the SUCEST data bank. Conserved regions corresponding to different metalloprotease domains and sequence motifs were identified in the reads to characterize each group of enzymes. At least four classes of sugarcane metalloproteases have been identified, i.e. matrix metalloproteases, zincins, inverzincins, and ATP-dependent metalloproteases. Each enzyme class was analyzed for its expression in different conditions and tissues.Metaloproteases exercem papéis importantes em muitos processos fisiológicos em mamíferos tais como migração celular, remodelamento tecidual e processamento de fatores de crescimento. Estas enzimas estão envolvidas também na pato-fisiologia de um grande número de doenças humanas como hipertensão e câncer. Muitas bactérias patogênicas dependem de proteases para infectar o hospedeiro. Diversas classes de metaloproteases foram descritas em seres humanos, bactérias, venenos de serpentes e insetos. No entanto, a presença e a caracterização de metaloproteases em plantas estão pouco descritas na literatura. Neste trabalho, foi pesquisada a biblioteca de cDNA de etiquetas de seqüências expressas da cana-de-açúcar (SUCEST) para identificar, por homologia com seqüências depositadas em outros bancos de dados, famílias gênicas de metaloproteases expressas em diferentes condições. Foram utilizadas seqüências protéicas de Arabidopis thaliana e Glycine max e seqüências de nucleotídeos de Sorghum bicolor. Regiões conservadas correspondentes aos diferentes domínios e motivos de seqüência de metaloproteases foram identificadas nos cDNAs de cana-de-açúcar para caracterizar cada grupo de enzimas. Pelo menos quatro classes de metaloproteases foram identificadas na cana-deaçúcar, a saber, metaloproteases de matriz extracelular, zincinas, inverzincinas e metaloproteases dependentes de ATP. Cada uma destas classes foi analisada quanto a sua expressão nas diferentes condições e tecidos utilizados na construção das bibliotecas de cDNA

Rat Skin Wound Healing Induced By Alternagin-c, A Disintegrin-like, Cys-rich Protein From Bothrops Alternatus Venom

Alternagin-C (ALT-C) is a disintegrin-like, Cys-rich protein isolated from Bothrops alternatus snake venom, which has been shown to induce in vivo angiogenesis. Therefore, this protein could be interesting as a new approach for tissue regeneration studies. Here the effects of ALT-C on fibroblasts and inflammatory cells, collagen type III and type I and TGF-α expression in a rat wounded skin model were studied. Thirty-five male Wistar rats (weight 270 ± 20 g) were divided into seven groups with five animals in each of the following groups: a control group which wounded animals received treatment with natrozol® gel only; ALT-C10, ALT-C60 and ALT-C100 groups of wounded animals that were treated with the same amount of gel containing 10, 60 and 100 ng of ALT-C, respectively. Animals were treated once a day with 20 μl of gel associated or not with ALT-C for 1, 3, 5 or 7 days. ALT-C treatment increased the fibroblast density, collagen deposition and accelerated the inflammatory process, mostly in the ALT-C60 group. These results indicate that ALT-C improves wound repair process in rat skin. Thus, ALT-C could be a candidate to the development of a novel therapeutic strategy for wounded skin repair. © 2011 The Authors. © 2011 Blackwell Publishing Ltd and Medicalhelplines.com Inc.83245252Singer, A.J., Clark, R.A., Cutaneous wound healing. (1999) N Engl J Med, 341, pp. 738-746Werner, S., Grose, R., Regulation of wound healing by growth factors and cytokines. (2003) Physiol Rev, 83, pp. 835-870Martin, P., Wound healing-aiming for perfect skin regeneration. (1997) Science, 276, pp. 75-81Clark, R.A., Basics of cutaneous wound repair. (1993) J Dermatol Surg Oncol, 19, pp. 693-706Schultz, G.S., Wysocki, A., Interactions between extracellular matrix and growth factors in wound healing. (2009) Wound Repair Regen, 17, pp. 153-162Tsahar, E., Moyer, J.D., Waterman, H., Barbacci, E.G., Bao, J., Levkowitz, G., Shelly, M., Yarden, Y., Pathogenic poxviruses reveal viral strategies to exploit the ErbB signaling network. (1998) EMBO J, 15, pp. 5948-5963Yang, X., Letterio, J.J., Lechleider, R.J., Chen, L., Hayman, R., Gu, H., Roberts, A.B., Deng, C., Targeted disruption of SMAD3 results in impaired mucosal immunity and diminished T cell responsiveness to TGF-beta. (1999) EMBO J, 18, pp. 1280-1291Strachan, L., Murison, J.G., Prestidge, R.L., Sleeman, M.A., Watson, J.D., Kumble, K.D., Cloning and biological activity of epigen, a novel member of the epidermal growth factor superfamily. (2001) J Biol Chem, 276, pp. 18265-18271Hashimoto, K., Regulation of keratinocyte function by growth factors. (2000) J Dermatol Sci, 24, pp. S46-S50Bennett, S.P., Griffiths, G.D., Schor, A.M., Leese, G.P., Schor, S.L., Growth factors in the treatment of diabetic foot ulcers. (2003) Br J Surg, 90, pp. 133-146Li, Y., Fan, J., Chen, M., Li, W., Woodley, D.T., Transforming growth factor-alpha: a major human serum factor that promotes human keratinocyte migration. (2006) J Invest Dermatol, 126, pp. 2096-2105Barrandon, Y., Green, H., Cell migration is essential for sustained growth of keratinocyte colonies: the roles of transforming growth factor-alpha and epidermal growth factor. (1987) Cell, 50, pp. 1131-1137Ellis, I.R., Schor, A.M., Schor, S.L., EGF AND TGF-α motogenic activities are mediated by the EGF receptor via distinct matrix-dependent mechanisms. (2007) Exp Cell Res, 313, pp. 732-741Kim, I., Mogford, J.E., Chao, J.D., Mustoe, T.A., Wound epithelialization deficits in the transforming growth factor-α knockout mouse. (2001) Wound Repair Regen, 9, pp. 386-390Mariano-Oliveira, A., Coelho, A.L., Terruggi, C.H., Selistre-de-Araújo, H.S., Barja-Fidalgo, C., De Freitas, M.S., Alternagin-C, a non-RGD-disintegrin, induces neutrophil migration via integrin signaling. (2003) Eur J Biochem, 270, pp. 4799-4808Selistre-de-Araujo, H.S., Cominetti, M.R., Terruggi, C.H., Mariano-Oliveira, A., De Freitas, M.S., Crepin, M., Figueiredo, C.C., Morandi, V., Alternagin-C, a disintegrin-like protein from the venom of Bothrops alternatus, modulates α2β1 integrin-mediated cell adhesion, migration and proliferation. (2005) Braz J Med Biol Res, 38, pp. 1505-1511Souza, D.H., Iemma, M.R., Ferreira, L.L., Faria, J.P., Oliva, M.L., Zingali, R.B., Niewiarowski, S., Selistre-de-Araujo, H.S., The disintegrin-like domain of the snake venom metalloprotease alternagin inhibits α2β1 integrin-mediated cell adhesion. (2000) Arch Biochem Biophys, 384, pp. 341-350Cominetti, M.R., Ribeiro, J.U., Fox, J.W., Selistre-de-Araujo, H.S., BaG, a new dimeric metalloproteinase/disintegrin from the Bothrops alternatus snake venom that interacts with α5β1 integrin. (2003) Arch Biochem Biophys, 416, pp. 171-179Ramos, O.H., Terruggi, C.H., Ribeiro, J.U., Cominetti, M.R., Figueiredo, C.C., Bérard, M., Crepin, M., Selistre-de-Araujo, H.S., Modulation of in vitro and in vivo angiogenesis by alternagin-C, a disintegrin-like protein from Bothrops alternatus snake venom and by a peptide derived from its sequence. (2007) Arch Biochem Biophys, 461, pp. 1-6Mesquita-Ferrari, R.A., Moraes, C.K., Micocci, K.C., Selistre-de-Araujo, H.S., ALT-C, a disintegrin-like Cys-rich protein from Bothrops alternatus, increases skeletal myoblast viability. (2009) J Venom Anim Toxins Incl Trop Disv, 15, pp. 325-339Durigan, J.L., Peviani, S.M., Russo, T.L., Delfino, G.B., Ribeiro, J.U., Cominetti, M.R., Selistre-de-Araujo, H.S., Salvini, T.F., Effects of alternagin-C from Bothrops alternatus on gene expression and activity of metalloproteinases in regenerating skeletal muscle. (2008) Toxicon, 52, pp. 687-694Sant'ana, E.M.C., Gouvêa, C.M.C.P., Nakaie, C.R., Selistre-de-Araújo, H.S., Angiogenesis and growth factor modulation induced by alternagin C, a snake venom disintegrin-like, cysteine-rich protein on a rat skin wound model. (2008) Arch Biochem Biophys, 479, pp. 20-27Junqueira, L.C.U., Cossermely, W., Brentani, R., Diferential staining of collagens type-I, Type-II and Type-III by picrosirius red and polarization microscopy. (1978) Arch Hist Jpn, 41, pp. 267-274Lapiere, C.M., Nusgens, B., Pierard, G.E., Interaction between collagen type I and type III in conditioning bundles organization. (1977) Connect Tissue Res, 5, pp. 21-29Robins, S.P., Milne, G., Duncan, A., Davies, C., Butt, R., Greiling, D., James, I.T., Increased skin collagen extractability and proportions of collagen type III are not normalized after 6 months healing of human excisional wounds. (2003) J Invest Dermatol, 121, pp. 267-272Witte, M., Barbul, A., General principles of wound healing. (1997) Surg Clin North Am, 77, pp. 509-528Terruggi, C.H.B., Cominetti, M.R., Bérard, M., Selistre-de-Araújo, H.S., The disintegrin alternagin-C modulates migration and viability of breast tumor cells. (2006) Rencontres en Toxinologie., pp. 201-8. , Toxines et cancer: LavoisierZigrino, P., Kamiguti, A.S., Eble, J., Drescher, C., Nischt, R., Fox, J.W., Mauch, C., The reprolysin jararhagin, a snake venom metalloproteinase, functions as a fibrillar collagen agonist involved in fibroblast cell adhesion and signaling. (2002) J Biol Chem, 277, pp. 40528-4053

Association of Bartonella spp bacteremia with Chagas cardiomyopathy, endocarditis and arrythmias in patients from South America

Infection with Bartonella spp may cause cardiac arrhythmias, myocarditis and endocarditis in humans. The aim of the present study was to evaluate a possible association between Bartonella spp bacteremia and endocarditis, arrhythmia and Chagas cardiomyopathy in patients from Brazil and Argentina. We screened for the presence of bacterial 16S rRNA in human blood by PCR using oligonucleotides to amplify a 185-bp bacterial DNA fragment. Blood samples were taken from four groups of subjects in Brazil and Argentina: i) control patients without clinical disease, ii) patients with negative blood-culture endocarditis, iii) patients with arrhythmias, and iv) patients with chronic Chagas cardiomyopathy. PCR products were analyzed on 1.5% agarose gel to visualize the 185-bp fragment and then sequenced to confirm the identity of DNA. Sixty of 148 patients (40.5%) with cardiac disease and 1 of 56 subjects (1.8%) from the control group presented positive PCR amplification for Bartonella spp, suggesting a positive association of the bacteria with these diseases. Separate analysis of the four groups showed that the risk of a Brazilian patient with endocarditis being infected with Bartonella was 22 times higher than in the controls. In arrhythmic patients, the prevalence of infection was 45 times higher when compared to the same controls and 40 times higher for patients with Chagas cardiomyopathy. To the best of our knowledge this is the first report of the association between Bartonella spp bacteremia and Chagas disease. The present data may be useful for epidemiological and prevention studies in Brazil and Argentina

Alternagin-C, a disintegrin-like protein from the venom of Bothrops alternatus, modulates a2ß1 integrin-mediated cell adhesion, migration and proliferation

The alpha2ß1 integrin is a major collagen receptor that plays an essential role in the adhesion of normal and tumor cells to the extracellular matrix. Alternagin-C (ALT-C), a disintegrin-like protein purified from the venom of the Brazilian snake Bothrops alternatus, competitively interacts with the alpha2ß1 integrin, thereby inhibiting collagen binding. When immobilized in plate wells, ALT-C supports the adhesion of fibroblasts as well as of human vein endothelial cells (HUVEC) and does not detach cells previously bound to collagen I. ALT-C is a strong inducer of HUVEC proliferation in vitro. Gene expression analysis was done using an Affimetrix HU-95A probe array with probe sets of ~10,000 human genes. In human fibroblasts growing on collagen-coated plates, ALT-C up-regulates the expression of several growth factors including vascular endothelial growth factor, as well as some cell cycle control genes. Up-regulation of the vascular endothelial growth factor gene and other growth factors could explain the positive effect on HUVEC proliferation. ALT-C also strongly activates protein kinase B phosphorylation, a signaling event involved in endothelial cell survival and angiogenesis. In human neutrophils, ALT-C has a potent chemotactic effect modulated by the intracellular signaling cascade characteristic of integrin-activated pathways. Thus, ALT-C acts as a survival factor, promoting adhesion, migration and endothelial cell proliferation after binding to alpha2ß1 integrin on the cell surface. The biological activities of ALT-C may be helpful as a therapeutic strategy in tissue regeneration as well as in the design of new therapeutic agents targeting alpha2ß1 integrin