23 research outputs found
Leptospirose humana: estudo sorológico de 29 anos em São Paulo, Brasil
A retrospective study of 9,335 cases of human leptospirosis in the state of São Paulo, Brazil, diagnosed between 1969 and 1997 showed that the disease is endemic throughout the state. Middle-aged adults, with a range of 20-39 years, were most frequently infected (32.40%). The mean annual incidence was 0.53 per 100,000 population and the disease was more frequent in males (87.0%). Cases occurred mainly in January to April each year. A peak was observed in 1991 and 1996 which rainfall average was 159.9 and 160.3, respectively. These data emphasize the potential public health importance of leptospirosis in the state of São Paulo, Brazil.Estudo retrospectivo com 9.335 casos de leptospirose humana no Estado de São Paulo, Brasil, diagnosticados entre 1969 e 1997 mostrou que a doença é endêmica no estado. Adultos com idade entre 20-39 anos foram os mais infectados (32,40%) sendo 87,0% dos casos do sexo masculino. A incidência anual média foi de 0,53 por uma população de 100.000. Casos ocorreram principalmente em janeiro a abril de cada ano. O maior número de casos foi observado em 1991 e 1996 com média pluviométrica de 159,9 e 160,3, respectivamente. Estes dados enfatizam a importância da leptospirose na saúde pública no Estado de São Paulo, Brasil
Characterization of Novel OmpA-Like Protein of Leptospira interrogans That Binds Extracellular Matrix Molecules and Plasminogen
Leptospira interrogans is the etiological agent of leptospirosis, a zoonotic disease of human and veterinary concern. The identification of novel proteins that mediate host-pathogen interactions is important for understanding the bacterial pathogenesis as well as to identify protective antigens that would help fight the disease. We describe in this work the cloning, expression, purification and characterization of three predicted leptospiral membrane proteins, LIC10258, LIC12880 (Lp30) and LIC12238. We have employed Escherichia coli BL21 (SI) strain as a host expression system. Recently, we have identified LIC12238 as a plasminogen (PLG)-binding receptor. We show now that Lp30 and rLIC10258 are also PLG-receptors of Leptospira, both exhibiting dose-dependent and saturating binding (KD, 68.8±25.2 nM and 167.39±60.1 nM, for rLIC10258 and rLIC12880, respectively). In addition, LIC10258, which is a novel OmpA-like protein, binds laminin and plasma fibronectin ECM molecules and hence, it was named Lsa66 (Leptospiral surface adhesin of 66 kDa). Binding of Lsa66 to ECM components was determined to be specific, dose-dependent and saturable, with a KD of 55.4±15.9 nM to laminin and of 290.8±11.8 nM to plasma fibronectin. Binding of the recombinant proteins to PLG or ECM components was assessed by using antibodies against each of the recombinant proteins obtained in mice and confirmed by monoclonal anti-polyhistidine antibodies. Lsa66 caused partial inhibition on leptospiral adherence to immobilized ECM and PLG. Moreover, this adhesin and rLIC12238 are recognized by antibodies in serum samples of confirmed leptospirosis cases. Thus, Lsa66 is a novel OmpA-like protein with dual activity that may promote the attachment of Leptospira to host tissues and may contribute to the leptospiral invasion. To our knowledge, this is the first leptospiral protein with ECM and PLG binding properties reported to date
Renal Involvement in Leptospirosis: The Effect of Glycolipoprotein on Renal Water Absorption
on vasopressin (Vp) action in the guinea pig inner medullary collecting duct (IMCD). Copenhageni, GLPc, n = 5); Group II, IMCD from normal guinea-pigs in the presence of GLPc (GLPc group, n = 54); Group III, IMCD from injected animals with GLPc ip (n = 8). (GLPp, non pathogenic, 250 µg) did not alter Vp action. In Group III, GLPc (250 µg) injected intraperitoneally produced a decrease of about 20% in IMCD Aquaporin 2 expression.The IMCD Pf decrease caused by GLP is evidence, at least in part, towards explaining the urinary concentrating incapacity observed in infected guinea-pigs
Aseptic meningitis caused by Leptospira spp diagnosed by polymerase chain reaction
Leptospirosis is a zoonotic disease caused by the pathogenic Leptospira
spp. The clinical presentations are diverse, ranging from
undifferentiated fever to fulminant disease including meningeal forms.
The neurological leptospirosis forms are usually neglected. The aim of
this study was to investigate leptospirosis as the cause of aseptic
meningitis using different diagnostic techniques including the
polymerase chain reaction (PCR). Thirty-nine cerebrospinal fluid (CSF)
samples from patients presenting with meningeal abnormalities,
predominance of lymphocytes and negative results by traditional
microbiological tests were processed by leptospiral culture,
anti-leptospiral antibody response and PCR. Leptospira spp DNA was
detected in 23 (58.97%) of the CSF samples. Anti-leptospiral antibodies
were found in 13 (33.33%) CSF samples. Twelve CSF samples were positive
by PCR assay and negative by microscopic agglutination test (MAT)
assay. Two CSF samples were positive by MAT and negative by PCR. The
positive and negative agreement between both tests was 11 and 14,
respectively. CSF samples from six cases of unknown diagnosis were
positive by PCR assay. Eight cases showed positive results using PCR
and MAT. Leptospirosis could be detected by PCR assay from the 3rd-26th
day after illness onset. The sensitivity of the PCR was assessed with
confirmed cases of leptospirosis (by MAT) and found to be 89.5%. All
CSFs were negative by culture. PCR was found to be a powerful tool for
diagnosing meningitis cases of leptospirosis. We recommend that it may
be used as a supplementary diagnostic tool, especially in the early
stages of the disease, when other diagnostic techniques such as
serology are not sensitive
Evaluation of nested polymerase chain reaction for the early detection of Leptospira spp. DNA in serum samples from patients with leptospirosis
In LipL32, the Major Leptospiral Lipoprotein, the C Terminus Is the Primary Immunogenic Domain and Mediates Interaction with Collagen IV and Plasma Fibronectin â–¿
LipL32 is the major leptospiral outer membrane lipoprotein expressed during infection and is the immunodominant antigen recognized during the humoral immune response to leptospirosis in humans. In this study, we investigated novel aspects of LipL32. In order to define the immunodominant domains(s) of the molecule, subfragments corresponding to the N-terminal, intermediate, and C-terminal portions of the LipL32 gene were cloned and the proteins were expressed and purified by metal affinity chromatography. Our immunoblot results indicate that the C-terminal and intermediate domains of LipL32 are recognized by sera of patients with laboratory-confirmed leptospirosis. An immunoglobulin M response was detected exclusively against the LipL32 C-terminal fragment in both the acute and convalescent phases of illness. We also evaluated the capacity of LipL32 to interact with extracellular matrix (ECM) components. Dose-dependent, specific binding of LipL32 to collagen type IV and plasma fibronectin was observed, and the binding capacity could be attributed to the C-terminal portion of this molecule. Both heparin and gelatin could inhibit LipL32 binding to fibronectin in a concentration-dependent manner, indicating that the 30-kDa heparin-binding and 45-kDa gelatin-binding domains of fibronectin are involved in this interaction. Taken together, our results provide evidence that the LipL32 C terminus is recognized early in the course of infection and is the domain responsible for mediating interaction with ECM proteins
Expression and characterization of HlyX hemolysin from Leptospira interrogans serovar Copenhageni: Potentiation of hemolytic activity by LipL32
The HlyX, a putative hemolysin identified from the Leptospira genomes, was cloned, expressed in Escherichia coli, purified, and its hemolytic activity was confirmed. Mouse polyclonal antiserum against the recombinant HlyX recognized HlyX-related antigens in a panel of Leptospira species extracts and it was also able to abolish the hemolytic activity of HlyX. A mixture of HlyX and LipL32, a known hemolysin from Leptospira, induced hemolysis in a synergistic way that was fully inhibited by antiserum against either protein. Moreover, sera from patients with leptospirosis also recognized the recombinant HlyX, showing that it is presented to the host immune system during Leptospira infection. © 2005 Elsevier Inc. All rights reserved.Fil: Hauk, Pricila. Governo do Estado de Sao Paulo. Secretaria da Saude. Instituto Butantan; Brasil. Universidade de Sao Paulo; BrasilFil: Negrotto, Soledad. Consejo Nacional de Investigaciones CientÃficas y Técnicas. Instituto de Medicina Experimental. Academia Nacional de Medicina de Buenos Aires. Instituto de Medicina Experimental; ArgentinaFil: Romero, Eliete Caló. Instituto Adolfo Lutz; BrasilFil: Vasconcellos, SÃlvio Arruda. Universidade de Sao Paulo; BrasilFil: Genovez, Margareth Élide. Centro de Pesquisa e Desenvolvimento de Sanidade Animal; BrasilFil: Ward, Richard John. Universidade de Sao Paulo; BrasilFil: Schattner, Mirta Ana. Consejo Nacional de Investigaciones CientÃficas y Técnicas. Instituto de Medicina Experimental. Academia Nacional de Medicina de Buenos Aires. Instituto de Medicina Experimental; ArgentinaFil: Gomez, Ricardo Martin. Consejo Nacional de Investigaciones CientÃficas y Técnicas. Centro CientÃfico Tecnológico Conicet - La Plata. Instituto de BiotecnologÃa y BiologÃa Molecular. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Instituto de BiotecnologÃa y BiologÃa Molecular; ArgentinaFil: Ho, Paulo Lee. Governo do Estado de Sao Paulo. Secretaria da Saude. Instituto Butantan; Brasil. Universidade de Sao Paulo; Brasi
Gene locus, protein name, gene bank reference sequence, features, gene conservation, sequence of the primers employed for DNA amplification, and molecular mass of expressed recombinant proteins.
1<p><a href="http://aeg.lbi.ic.unicamp.br/world/lic/" target="_blank">http://aeg.lbi.ic.unicamp.br/world/lic/</a>;</p>2<p><a href="http://www.ncbi.nlm.nih.gov/protein/" target="_blank">http://www.ncbi.nlm.nih.gov/protein/</a>;</p>3<p><a href="http://www.ncbi.nlm.nih.gov/blast/Blast.cgi/" target="_blank">http://www.ncbi.nlm.nih.gov/blast/Blast.cgi/</a>. This work, Lai: <i>L. interrogans</i> serovar Lai; LBH: <i>L. borgpetersenii</i> serovar Hardjo-bovis; LBP: <i>L. biflexa</i> serovar Patoc.</p