RESUMEN
In this study, we associated the restriction modification (RM) tests to the polymerase chain reaction (PCR) detection of molecular markers (SCCmec III, seh, agr II-SCCmec IV, and lukSF) for revealing the main methicillin-resistant Staphylococcus aureus (MRSA) clones circulating in Brazil. This simple and rapid approach allowed a precise classification of the MRSA analyzed when compared with pulsed-field gel electrophoresis (PFGE) data.
Asunto(s)
Enzimas de Restricción-Modificación del ADN , Staphylococcus aureus Resistente a Meticilina/clasificación , Staphylococcus aureus Resistente a Meticilina/aislamiento & purificación , Tipificación Molecular/métodos , Reacción en Cadena de la Polimerasa/métodos , Infecciones Estafilocócicas/microbiología , Brasil , Análisis por Conglomerados , Electroforesis en Gel de Campo Pulsado , Humanos , Staphylococcus aureus Resistente a Meticilina/genéticaRESUMEN
OBJECTIVES: Sepsis is a "life-threatening organ dysfunction caused by a dysregulated host response to infection", which is identified by a >2 point increase from patient baseline in the Sequential Organ Failure Assessment score (SOFAs). The prevalence and outcome of patients with sepsis has been mainly assessed in ICU patients, while few data are available for patients admitted to internal medicine wards. Our purpose was to evaluate the prevalence and the clinical outcome of patients with sepsis in an internal medicine-ward. DESIGN: This is a single-center retrospective observational study evaluating all patients admitted over a 2-month period (October and Novembre 2015) in the internal medicine ward of the San Giovanni di Dio Hospital in Florence. Patients with clinical and/or instrumental signs of bacterial infection were evaluated with SOFAs and divided into patients with and without sepsis. RESULTS: 635 patients were evaluated, and 279 of them (43.9%) were diagnosed with a bacterial infection. The diagnosis of sepsis was made in 93 patients (14.6%). In-hospital mortality and transfer to ICU were observed in 16% of patients with sepsis and in 2.5% of patients without sepsis (p<0.0001). A SOFAs value <2 had a negative predic-tive value of 97.5%, and increasing values of SOFAs were associated with a worse prognosis. CONCLUSIONS: The results suggest that: a) a high proportion of patients hospitalized in an internal medicine ward are affected by sepsis; b) these patients are burdened with high in-hospital mortality or transfer to ICU; c) SOFA score has a high prognostic power.
Asunto(s)
Unidades de Cuidados Intensivos/estadística & datos numéricos , Sepsis/mortalidad , Anciano , Anciano de 80 o más Años , Femenino , Mortalidad Hospitalaria , Hospitales/estadística & datos numéricos , Humanos , Masculino , Persona de Mediana Edad , Puntuaciones en la Disfunción de Órganos , Prevalencia , Pronóstico , Estudios RetrospectivosRESUMEN
Endothelial cell receptors for the angiostatic chemokines IFN-gamma-inducible protein of 10 kDa (IP-10) and monokine induced by IFN-gamma (Mig) have not yet been identified, and the mechanisms responsible for the effects of these chemokines on angiogenesis are still unclear. IP-10 and Mig share a common functional receptor on activated T lymphocytes, named CXC chemokine receptor 3 (CXCR3). Using in situ hybridization and immunohistochemistry, we show that CXCR3 is expressed by a small percentage of microvascular endothelial cells in several human normal and pathological tissues. Primary cultures of human microvascular endothelial cells (HMVECs) likewise express CXCR3, although this expression is limited to the S/G2-M phase of their cell cycle. Both IP-10 and Mig, as well as the IFN-gamma-inducible T-cell alpha chemoattractant (I-TAC), which all share high-affinity binding for CXCR3, block HMVEC proliferation in vitro, an effect that can be inhibited by an anti-CXCR3 antibody. These data provide definitive evidence of CXCR3 expression by HMVEC and open new avenues for therapeutic interventions in all conditions in which an angiostatic effect may be beneficial.
Asunto(s)
Endotelio Vascular/metabolismo , Péptidos y Proteínas de Señalización Intercelular , Receptores de Quimiocina/genética , Receptores de Quimiocina/metabolismo , Inhibidores de la Angiogénesis/farmacología , Ciclo Celular , División Celular/efectos de los fármacos , Células Cultivadas , Quimiocina CXCL10 , Quimiocina CXCL11 , Quimiocina CXCL9 , Quimiocinas CXC/farmacología , Endotelio Vascular/citología , Endotelio Vascular/efectos de los fármacos , Expresión Génica , Humanos , Neovascularización Fisiológica , ARN Mensajero/genética , ARN Mensajero/metabolismo , Receptores CXCR3 , Receptores de Quimiocina/antagonistas & inhibidores , Distribución TisularRESUMEN
The preferential association of some chemokine receptors with human Th1 or Th2 cells has recently been reported. In this study, the expression of CCR3, CCR5, CXCR3, and CXCR4 were analyzed by flow cytometry in three distinct in vitro models of Th1/Th2 polarization, activated naive and memory T cells, and T-cell clones, in which the intracellular synthesis of interferon-gamma (IFN-gamma) and interleukin-4 (IL-4) and the surface expression of CD30 and LAG-3 were also assessed. Moreover, by using immunohistochemistry the in vivo expression of CCR3, CCR5, CXCR3, and CXCR4 was examined in the gut of patients suffering from Crohn's disease, a Th1-dominated disorder, and in the skin of patients suffering from systemic sclerosis, a Th2-dominated disorder. CCR5 and LAG-3 exhibited the same pathway of Th1 association, whereas CXCR3 did not discriminate between Th1- and Th2-dominated responses. On the other hand, CCR3 was found only occasionally in a small proportion of allergen-specific memory T cells with Th2/ThO profile of cytokine production in vitro. However, it was neither seen in Th2-polarized activated naive T cells nor in established Th2 clones and could be detected in vivo only on non-T cells. Finally, whereas CXCR4 expression was not limited to Th2 cells in vivo, it was markedly up-regulated by IL-4 and down-regulated by IFN-gamma in vitro. Thus, the results of this study confirm the existence of flexible programs of chemokine receptor expression during the development of Th1 and Th2 cells. However, caution is advised in interpreting these receptors as surrogate markers of a given type of effector response.
Asunto(s)
Receptores de Quimiocina/biosíntesis , Células TH1/metabolismo , Células Th2/metabolismo , Células Clonales , Enfermedad de Crohn/inmunología , Enfermedad de Crohn/metabolismo , Regulación hacia Abajo/efectos de los fármacos , Citometría de Flujo , Humanos , Memoria Inmunológica , Interferón gamma/biosíntesis , Interferón gamma/farmacología , Interferón gamma/fisiología , Interleucina-12/farmacología , Interleucina-4/biosíntesis , Interleucina-4/farmacología , Interleucina-4/fisiología , Activación de Linfocitos , Especificidad de Órganos , Regulación hacia Arriba/efectos de los fármacosRESUMEN
Biofilm formed by Staphylococcus aureus is considered an important virulence trait in the pathogenesis of infections associated with implantable medical devices. Gene expression analyses are important strategies for determining the mechanisms involved in production and regulation of biofilm. Obtaining intact RNA preparations is the first and most critical step for these studies. In this article, we describe an optimized protocol for obtaining total RNA from sessile cells of S. aureus using the RNeasy Mini Kit. This method essentially consists of a few steps, as follows: 1) addition of acetone-ethanol to sessile cells, 2) lysis with lysostaphin at 37°C/10 min, 3) vigorous mixing, 4) three cycles of freezing and thawing, and 5) purification of the lysate in the RNeasy column. This simple pre-kit procedure yields high-quality total RNA from planktonic and sessile cells of S. aureus.
Asunto(s)
Técnicas Bacteriológicas/normas , Biopelículas/crecimiento & desarrollo , ARN Bacteriano/aislamiento & purificación , Staphylococcus aureus/genética , Técnicas Bacteriológicas/métodos , Electroforesis en Gel de Agar , Proteínas Hemolisinas/metabolismo , Staphylococcus aureus Resistente a Meticilina/genética , Staphylococcus aureus Resistente a Meticilina/aislamiento & purificación , Control de Calidad , Reacción en Cadena en Tiempo Real de la Polimerasa , Transcripción Reversa , Staphylococcus aureus/fisiologíaRESUMEN
The gas-phase basicities of serine and dipeptides containing amino acid residues of serine and glycine were determined by proton transfer reactions in a Fourier transform ion cyclotron resonance mass spectrometer. The gas-phase basicity (GB) of L-serine was found to be 205.9 kcal/mol, with addition of a hydroxymethyl group (-CH2OH) increasing the basicity by 4.5 kcal/mol relative to the simplest amino acid glycine (GB = 201.4 kcal/mol). This is attributed to a combination of intramolecular hydrogen bonding, induction, and symmetry effects. For the dipeptides, addition of a hydroxymethyl group does not result in a large increase in basicity relative to the basicity of glycylglycine (GB = 208.0 kcal/mol). The gas-phase basicities determined for glycyl-L-serine, L-serylglycine, and L-sery-L-serine are 209.3,210.6, and 210.9 kcal/mol, respectively. In comparison to glycylglycine, addition of the hydroxymethyl group at the N terminus has a greater impact on basicity than its placement at the C terminus. These data suggest that the protonation site for these dipeptides is the N-terminal amino nitrogen.
RESUMEN
The genetic effects of nitrilotriacetic acid (NTA) and ethylenedinitrilotetraacetic acid (EDTA), two widely used chelating agents, were investigated by using a somatic mutation and recombination test (SMART) after treatment of larvae and the FIX test for aneuploidy after treatment of adult female Drosophila melanogaster. Chloral hydrate (CH) and 5-fluorodeoxyuridine (FdUr) were used as positive controls. Effectively absorbed amounts of the test compounds assayed in Drosophila were estimated at the single fly level by a method using 3H-leucine. NTA and EDTA were also assayed in tests for aneuploidy based on chromosome counting in mouse germ and somatic cells. We previously showed that NTA was able to induce aneuploidy (chromosomal gain) in the germ cells of both Drosophila and the mouse when tested at the exposure levels of 5 x 10(-2) M and 275 mg per kg body weight, respectively [Costa et al., Environ Mol Mutagen 12:397-407, 1988]. In the present experiments, EDTA was assayed at 2.5 x 10(-2) M and 7.5 x 10(-3) M in the FIX test adopting a three-stage brooding scheme. Significant increases (with respect to controls) in chromosomal loss were observed in the second brood and in the combined three-brood total for both exposure levels of EDTA. In the SMART test, treatments with EDTA in the same exposure range produced negative results over all end-points, whereas significant increases in the frequency of small single spots (possibly due to aneuploidy) were produced by NTA 5 x 10(-2) M. In the cytogenetic assays for aneuploidy both in the germ and somatic cells of the mouse, negative results were also obtained following the i.p. administration of 93 and 186 mg EDTA per kg b.w. The previously observed induction of germ cell aneuploidy by NTA (275 mg per kg b.w.) was confirmed in the present experiments on a different strain of mice. NTA (138-275 mg per kg b.w.) did not induce aneuploidy in somatic cells of the mouse [Russo et al., Mutat Res 226: 111-114, 1989], however. These results are compared and discussed with reference to the characteristics of the different test systems used and to the different chelating properties of NTA and EDTA.
Asunto(s)
Acetatos/toxicidad , Aneuploidia , Ácido Edético/toxicidad , Mutación , Ácido Nitrilotriacético/toxicidad , Animales , Línea Celular , Drosophila melanogaster/efectos de los fármacos , Drosophila melanogaster/genética , Femenino , Células Germinativas/efectos de los fármacos , Masculino , Ratones , Recombinación GenéticaRESUMEN
The frequencies of micronuclei induced by ZnCl2 and detected on the gill tissue of the marine mussel Mytilus galloprovincialis with different staining techniques (acridine orange, gallocyanin chromallum, Feulgen, Giemsa) were compared. At least in the used system, the Feulgen and gallocyanin chromallum methods gave a frequency of micronuclei significantly lower than that obtained with the acridine orange and Giemsa techniques. No significant difference between the frequencies obtained with acridine orange and Giemsa was shown. So, though the acridine orange is surely the method which provides the more reliable data, in environmental screening works the Giemsa technique may be more suitable for its simplicity.
Asunto(s)
Bivalvos/efectos de los fármacos , Cloruros/farmacología , Compuestos de Zinc , Zinc/farmacología , Animales , Núcleo Celular/efectos de los fármacos , Pruebas de Mutagenicidad , Coloración y EtiquetadoRESUMEN
The genotoxicity of a chelating agent, the trisodium salt of nitrilotriacetic acid (NTA), was assessed in a somatic mutation and recombination test (SMART) in Drosophila melanogaster employing the wing hair markers mwh and flr3. The experiments were performed in parallel in two different laboratories (Padua, Italy and Schwerzenbach, Switzerland). The effectively absorbed doses of NTA, which was administered by feeding to larvae, were determined by a sensitive method employing [3H]leucine which allowed individual consumption levels to be measured. The particular pattern of clone induction produced by this compound suggests that NTA is active in inducing mitotic recombination and possibly aneuploidy in somatic cells of Drosophila. This is discussed in relation to the data present in the literature regarding the genotoxicity of NTA in a variety of experimental systems.
Asunto(s)
Quelantes/toxicidad , Ácido Nitrilotriacético/toxicidad , Animales , Drosophila melanogaster/efectos de los fármacos , Drosophila melanogaster/genética , Ingestión de Alimentos , Frecuencia de los Genes , Marcadores Genéticos , Pruebas de Mutagenicidad , Mutación , Sensibilidad y EspecificidadRESUMEN
A case of spontaneous dorso- lumbar spinal epidural hematoma during pregnancy is reported. The hematoma was removed 8 hours after the onset of paraplegia, and there was no evidence of vascular malformation. The motor deficit remained unchanged post-operatively. The etiology, clinical findings and the value of early laminectomy are discussed.
Asunto(s)
Hematoma Epidural Craneal , Complicaciones del Embarazo , Adulto , Femenino , Hematoma Epidural Craneal/complicaciones , Hematoma Epidural Craneal/cirugía , Humanos , Embarazo , Complicaciones del Embarazo/cirugía , Compresión de la Médula Espinal/etiologíaRESUMEN
In this study, genotyping techniques including staphylococcal chromosomal cassette mec (SCCmec) typing, pulsed-field gel electrophoresis (PFGE), multilocus sequence typing (MLST) and restriction-modification tests were used to compare the molecular characteristics of methicillin-resistant Staphylococcus aureus (MRSA) isolates recovered at two times within a 10-year interval (1998 and 2008) from a tertiary Brazilian hospital. In addition, the antimicrobial susceptibility profiles were analyzed. All 48 MRSA isolates from 1998 and 85.7% from 2008 (48/56 isolates) displayed multidrug-resistance phenotypes and SCCmec III. All but one of the 13 representative SCCmec III isolates belonged to CC8 and had PFGE patterns similar to that of the BMB9393 strain (Brazilian epidemic clone of MRSA; BEC). In 2008, we found an increased susceptibility to rifampicin and chloramphenicol among the SCCmec III isolates. In addition, we detected the entrance of diverse international MRSA lineages susceptible to trimethoprim-sulfamethoxazole (SXT), almost all belonging to CC5. These non-SCCmec III isolates were related to the USA 300 (ST8-SCCmec IV; PFGE-type B), USA 800 (ST5-SCCmec IV; subtype D1), USA 100 (ST5-SCCmec II; subtype D2), and EMRSA-3/Cordobes (ST5-SCCmec I, type C) clones. To the best of our knowledge, this is the first report of the emergence of isolates genetically related to the EMRSA-3/Cordobes clone in southeast Brazil. In this regard, these isolates were the most common non-SCCmec III MRSA in our institution, accounting for 8.9% of all isolates recovered in 2008. Thus, despite the supremacy of BEC isolates in our country, significant changes may occur in local MRSA epidemiology, with possible consequences for the rationality of MRSA empiric therapy.
Asunto(s)
Antibacterianos/farmacología , Staphylococcus aureus Resistente a Meticilina/efectos de los fármacos , Combinación Trimetoprim y Sulfametoxazol/farmacología , Brasil , ADN Bacteriano/genética , Electroforesis en Gel de Campo Pulsado , Genotipo , Humanos , Staphylococcus aureus Resistente a Meticilina/genética , Staphylococcus aureus Resistente a Meticilina/aislamiento & purificación , Pruebas de Sensibilidad Microbiana , Tipificación de Secuencias Multilocus , Fenotipo , Factores de TiempoRESUMEN
PURPOSE: This qualitative study explored barriers to implementing evidence-based practices (EBPs) in community-based addiction treatment organizations (CBOs) by comparing staff descriptions of barriers for four EBPs: Motivational Interviewing (MI), Adolescent Community Reinforcement Approach (A-CRA), Assertive Community Treatment (ACT), and Cognitive-behavioral Therapy (CBT). METHODS: The CBOs received CSAT/SAMHSA funding from 2003 to 2008 to deliver services using EBPs. Phone interview responses from 172 CBO staff directly involved in EBP implementation were analyzed using content analysis, a method for making inferences and developing themes from the systematic review of participant narratives (Berelson, 1952). RESULTS: Staff described different types of barriers to implementing each EBP. For MI, the majority of barriers involved staff resistance or organizational setting. For A-CRA, the majority of barriers involved specific characteristics of the EBP or client resistance. For CBT, the majority of barriers were associated with client resistance, and for ACT, the majority of barriers were associated with resources. DISCUSSION: EBP designers, policy makers who support EBP dissemination and funders should include explicit strategies to address such barriers. Addiction programs proposing to use specific EBPs must consider whether their programs have the organizational capacity and community capacity to meet the demands of the EBP selected.
Asunto(s)
Terapia Cognitivo-Conductual , Medicina Basada en la Evidencia/métodos , Accesibilidad a los Servicios de Salud , Necesidades y Demandas de Servicios de Salud , Motivación , Trastornos Relacionados con Sustancias/terapia , Adolescente , Conducta del Adolescente , Asertividad , Servicios Comunitarios de Salud Mental , Política de Salud , Disparidades en el Estado de Salud , Humanos , Entrevista Psicológica , National Institute on Drug Abuse (U.S.) , Proyectos Piloto , Investigación Cualitativa , Centros de Tratamiento de Abuso de Sustancias , Grabación en Cinta , Estados Unidos , United States Substance Abuse and Mental Health Services AdministrationRESUMEN
Biofilm formed by Staphylococcus aureus is considered an important virulence trait in the pathogenesis of infections associated with implantable medical devices. Gene expression analyses are important strategies for determining the mechanisms involved in production and regulation of biofilm. Obtaining intact RNA preparations is the first and most critical step for these studies. In this article, we describe an optimized protocol for obtaining total RNA from sessile cells of S. aureus using the RNeasy Mini Kit. This method essentially consists of a few steps, as follows: 1) addition of acetone-ethanol to sessile cells, 2) lysis with lysostaphin at 37°C/10 min, 3) vigorous mixing, 4) three cycles of freezing and thawing, and 5) purification of the lysate in the RNeasy column. This simple pre-kit procedure yields high-quality total RNA from planktonic and sessile cells of S. aureus.
Asunto(s)
Técnicas Bacteriológicas/normas , Biopelículas/crecimiento & desarrollo , ARN Bacteriano/aislamiento & purificación , Staphylococcus aureus/genética , Técnicas Bacteriológicas/métodos , Electroforesis en Gel de Agar , Proteínas Hemolisinas/metabolismo , Staphylococcus aureus Resistente a Meticilina/genética , Staphylococcus aureus Resistente a Meticilina/aislamiento & purificación , Control de Calidad , Reacción en Cadena en Tiempo Real de la Polimerasa , Transcripción Reversa , Staphylococcus aureus/fisiologíaRESUMEN
In this study, genotyping techniques including staphylococcal chromosomal cassette mec (SCCmec) typing, pulsed-field gel electrophoresis (PFGE), multilocus sequence typing (MLST) and restriction-modification tests were used to compare the molecular characteristics of methicillin-resistant Staphylococcus aureus (MRSA) isolates recovered at two times within a 10-year interval (1998 and 2008) from a tertiary Brazilian hospital. In addition, the antimicrobial susceptibility profiles were analyzed. All 48 MRSA isolates from 1998 and 85.7% from 2008 (48/56 isolates) displayed multidrug-resistance phenotypes and SCCmec III. All but one of the 13 representative SCCmec III isolates belonged to CC8 and had PFGE patterns similar to that of the BMB9393 strain (Brazilian epidemic clone of MRSA; BEC). In 2008, we found an increased susceptibility to rifampicin and chloramphenicol among the SCCmec III isolates. In addition, we detected the entrance of diverse international MRSA lineages susceptible to trimethoprim-sulfamethoxazole (SXT), almost all belonging to CC5. These non-SCCmec III isolates were related to the USA 300 (ST8-SCCmec IV; PFGE-type B), USA 800 (ST5-SCCmec IV; subtype D1), USA 100 (ST5-SCCmec II; subtype D2), and EMRSA-3/Cordobes (ST5-SCCmec I, type C) clones. To the best of our knowledge, this is the first report of the emergence of isolates genetically related to the EMRSA-3/Cordobes clone in southeast Brazil. In this regard, these isolates were the most common non-SCCmec III MRSA in our institution, accounting for 8.9% of all isolates recovered in 2008. Thus, despite the supremacy of BEC isolates in our country, significant changes may occur in local MRSA epidemiology, with possible consequences for the rationality of MRSA empiric therapy.
Asunto(s)
Humanos , Antibacterianos/farmacología , Staphylococcus aureus Resistente a Meticilina/efectos de los fármacos , Combinación Trimetoprim y Sulfametoxazol/farmacología , Brasil , ADN Bacteriano/genética , Electroforesis en Gel de Campo Pulsado , Genotipo , Pruebas de Sensibilidad Microbiana , Tipificación de Secuencias Multilocus , Staphylococcus aureus Resistente a Meticilina/genética , Staphylococcus aureus Resistente a Meticilina/aislamiento & purificación , Fenotipo , Factores de TiempoRESUMEN
BACKGROUND: The aim of this study was to investigate whether human mast cells express functional active CCR3 receptors, which are activated by CC chemokines. These ligands include the CCR3-selective chemokines eotaxin and eotaxin-2 and the more promiscuous CC chemokines, MCP-4, MCP-3, MCP-2 and RANTES. METHODS: Immunohistochemical analysis was performed on skin, gut and lung specimens. Double immunostaining was performed with anti-CCR3 and antitryptase, and anti-CCR3 and antichymase antibody (Ab) by using the avidin-biotin-peroxidase system with two different substrates. Mast cells were isolated and purified from human lung parenchyma (HLMC) by countercurrent elutriation followed by discontinuous Percoll density gradient. Flow-cytometric analysis of HLMC surface CCR3 expression was performed with the monoclonal Ab anti-CCR3 (7B11). Functional activation of HLMC was verified by the ability of cells to release histamine and/or migrate in response to eotaxin. RESULTS: High percentages (>70%) of tryptase-positive cells showing CCR3 expression were found in the skin and in the intestinal submucosa, whereas much lower percentages (< or = 20%) were found in the intestinal mucosa and in the lung interstitium. Eotaxin (1-100 nM) neither induced histamine release from HLMC nor enhanced anti-IgE-induced histamine release. In contrast, eotaxin (10-100 nM) and RANTES (10-100 nM) induced HLMC chemotaxis in vitro. Preincubation of HLMC with antibody anti-CCR3 (5 microg/ml) before loading into the chemotaxis chamber abrogated chemotaxis elicited by eotaxin. Double immunostaining with anti-CCR3 and anti-chymase antibody showed that the vast majority of CCR3-expressing mast cells in the various human tissues examined were tryptase-chymase double-positive. CONCLUSIONS: These results indicate that CCR3 is expressed on human mast cells and that these cells are attracted by CCR3-binding chemokines.
Asunto(s)
Quimiocinas CC , Mastocitos/metabolismo , Receptores de Quimiocina/metabolismo , Células Cultivadas , Quimiocina CCL11 , Quimiocina CCL5/farmacología , Quimiotaxis , Citocinas/farmacología , Humanos , Mucosa Intestinal/metabolismo , Pulmón/citología , Pulmón/metabolismo , Mastocitos/efectos de los fármacos , Mastocitos/fisiología , Receptores CCR3 , Piel/metabolismoRESUMEN
Eosinophils, basophils, and Th2 cells express the chemokine receptor CCR3, which binds eotaxin, RANTES, and some other chemokines. Using immunohistochemistry and flow cytometry, we demonstrate that CCR3 is also expressed by a variable proportion of human mast cells in gut, skin, and lung tissue. By contrast, with the same anti-CCR3 antibody (B711), CCR3 was poorly if at all detectable on human Th2 cells in vitro and in vivo. Eotaxin neither induced histamine release from purified human mast cells nor increased anti-IgE-stimulated histamine secretion. However, both eotaxin and RANTES elicited mast cell migration in vitro with a similar efficacy. High percentages of CCR3-expressing mast cells were present in the skin and in the intestinal submucosa; much lower percentages were found in the intestinal mucosa and in lung interstitium. Double immunostaining with anti-CCR3 and anti-chymase antibody showed that the vast majority of CCR3-expressing mast cells in the various tissues examined were tryptase-chymase double-positive. Therefore, tryptase-chymase double-positive mast cells express CCR3 and are attracted by CCR3-binding chemokines, eotaxin, and RANTES. Our findings indicate that these chemokines may play an important role in the differentiation and/or migration of this mast cell subset in connective tissues, as well as in sites of allergic inflammation.
Asunto(s)
Quimiocinas/metabolismo , Mastocitos/enzimología , Receptores de Quimiocina/biosíntesis , Ribonucleasas , Serina Endopeptidasas/biosíntesis , Proteínas Sanguíneas/biosíntesis , Células Cultivadas , Quimiotaxis , Quimasas , Proteínas en los Gránulos del Eosinófilo , Eosinófilos/metabolismo , Citometría de Flujo , Humanos , Inmunohistoquímica , Mastocitos/citología , Especificidad de Órganos , Receptores CCR3 , Células Th2/citología , Células Th2/enzimología , TriptasasRESUMEN
The mechanisms responsible for mesangial cell proliferation in proliferative glomerulonephritis are only partially understood. This article reports the results of an immunohistochemical study showing high expression of the chemokine receptor CXCR3 by mesangial cells of patients with IgA nephropathy, membranoproliferative glomerulonephritis, or rapidly progressive glomerulonephritis. CXCR3 was also detectable by flow cytometry in cultured human mesangial cells, in which it appeared to be functionally active, as determined by the ability of its ligand, the (interferon-gamma)-inducible protein of 10 kD (IP-10) to induce intracellular Ca2+ influx. Both IP-10 and the monokine induced by interferon-gamma (Mig) were also effective in inducing proliferation of human mesangial cells. These data suggest that in patients with proliferative glomerulonephritis, the chemokines IP-10 and/or Mig not only may act as chemoattractants for infiltrating mononuclear cells in the inflamed tissue, but also may directly induce the proliferation of mesangial cells.
Asunto(s)
Quimiocinas CXC/metabolismo , Glomerulonefritis/inmunología , Receptores de Quimiocina/metabolismo , Adulto , Anciano , Calcio/metabolismo , Estudios de Casos y Controles , División Celular , Células Cultivadas , Quimiocina CXCL10 , Glomerulonefritis/metabolismo , Glomerulonefritis/patología , Glomerulonefritis por IGA/inmunología , Glomerulonefritis por IGA/metabolismo , Glomerulonefritis por IGA/patología , Glomerulonefritis Membranoproliferativa/inmunología , Glomerulonefritis Membranoproliferativa/metabolismo , Glomerulonefritis Membranoproliferativa/patología , Humanos , Riñón/inmunología , Riñón/metabolismo , Riñón/patología , Persona de Mediana Edad , Receptores CXCR3RESUMEN
The chemoattractant activity of macrophage-derived chemokine (MDC), EBI1-ligand chemokine (ELC), and secondary lymphoid tissue chemokine (SLC) on human thymocytes was analyzed. Both ELC and SLC caused the accumulation of CD4+CD8- or CD4-CD8+ CD45RA+ thymocytes showing high CD3 expression. By contrast, a remarkable proportion of MDC-responsive thymocytes were CD4+CD8+ cells exhibiting reduced levels of CD8 or CD4+CD8- cells showing CD3 and CD45R0, but not CD45RA. MDC-responsive thymocyte suspensions were enriched in cells expressing the MDC receptor, CCR4, selectively localized to the medulla, and in CD30+ cells, whereas ELC-responsive thymocytes never expressed CD30. Reactivity to both MDC and ELC was localized to cells of the medullary areas, but never in the cortex. Double immunostaining showed no reactivity for either MDC or ELC by T cells, macrophages, or mature dendritic cells, whereas many medullary epithelial cells were reactive to MDC or ELC. However, MDC reactivity was consistently localized to the outer wall of Hassal's corpuscles, whereas ELC reactivity was often found in cells surrounding medullary vessels, but not in Hassal's corpuscles. Moreover, while most MDC-producing cells also stained positive for CD30L, this molecule was never found on ELC-producing cells. We suggest therefore that CD30L-expressing MDC-producing medullary epithelial cells attract CCR4-expressing thymocytes, thus favoring the CD30/CD30L interaction, and therefore the apoptosis, of cells that are induced to express CD30 by autoantigen activation. By contrast, ELC production by CD30L-lacking medullary epithelial cells may induce the migration into periphery of mature thymocytes that have survived the process of negative selection.
Asunto(s)
Quimiocinas CC/fisiología , Células Epiteliales/inmunología , Células Epiteliales/metabolismo , Subgrupos de Linfocitos T/citología , Subgrupos de Linfocitos T/inmunología , Timo/inmunología , Diferenciación Celular/inmunología , Quimiocina CCL19 , Quimiocina CCL21 , Quimiocina CCL22 , Quimiocinas CC/biosíntesis , Quimiotaxis de Leucocito/inmunología , Preescolar , Células Epiteliales/clasificación , Células Epiteliales/citología , Humanos , Lactante , Recién Nacido , Antígeno Ki-1/biosíntesis , Antígenos Comunes de Leucocito/biosíntesis , Ligandos , Timo/citología , Timo/metabolismoRESUMEN
Strong reactivity for interferon-inducible protein 10 (IP-10), monokine induced by interferon gamma (Mig), and interferon-inducible T-cell alpha chemoattractant (I-TAC) was found in epithelial cells mainly localized to the medulla of postnatal human thymus. The CXC chemokine receptor common to the 3 chemokines (CXCR3) was also preferentially expressed in medullary areas of the same thymuses and appeared to be a property of 4 distinct populations: CD3+ T-cell receptor (TCR) alphabeta+ CD8+ single-positive (SP) T cells, TCRgammadelta+ T cells, natural killer (NK)-type cells, and a small subset of CD3+(low) CD4+ CD8+ TCRalphabeta+ double-positive (DP) T cells. IP-10, Mig, and I-TAC showed chemoattractant activity for TCRalphabeta+ CD8+ SP T cells, TCRgammadelta+ T cells, and NK-type cells, suggesting their role in the migration of different subsets of mature thymocytes during human thymus lymphopoiesis.
Asunto(s)
Quimiocinas CXC/inmunología , Quimiotaxis de Leucocito , Péptidos y Proteínas de Señalización Intercelular , Células Asesinas Naturales/inmunología , Linfocitos T/inmunología , Timo/metabolismo , Linfocitos T CD8-positivos/inmunología , Células Cultivadas , Quimiocina CXCL10 , Quimiocina CXCL11 , Quimiocina CXCL9 , Quimiocinas CXC/biosíntesis , Quimiocinas CXC/genética , Células Epiteliales/inmunología , Humanos , Lactante , Recién Nacido , Subgrupos Linfocitarios/clasificación , ARN Mensajero/biosíntesis , Receptores de Antígenos de Linfocitos T alfa-beta/inmunología , Receptores de Antígenos de Linfocitos T gamma-delta/inmunología , Receptores CXCR3 , Receptores de Quimiocina/biosíntesis , Timo/citología , Timo/inmunologíaRESUMEN
Registro de caso de paciente no oitavo mes de gestacao que desenvolveu hematoma epidural espinal espontaneo dorsolombar. A gravidez, determinando aumento da pressao intra-abdominal e, como consequencia, aumento da pressao venosa no plexo epidural, poderia ter sido o fator desencadeante no hematoma. A paciente foi submetida a cirurgia precocemente, porem nao apresentou recuperacao do deficit sensitivo-motor.Sao discutidos aspectos clinicos, do tratamento cirurgico, da evolucao e da etiologia dos hematomas epidurais espinais espontaneos