RESUMEN
Toxoplasma gondii is a prevalent parasite of mammals and birds including up to 30% of humans world-wide. Primary infection of immunocompetent hosts leads to a robust cell-mediated immune response, which controls but does not clear the infection, thus enabling long-term parasite persistence in brain and muscle tissues. Chronic toxoplasmosis in mice is associated with resistance to heterologous pathogens and this has been related to increased numbers of inflammatory monocytes. Here we have analyzed whether chronic T. gondii infection impacts the subset distribution and the phenotype of peripheral human monocytes in vivo and their responses to parasite infection in vitro. CD14+ monocytes from T. gondii-seropositive blood donors expressed significantly less FcγRIII (CD16) than those from seronegative controls, but they did not show a shift in the distribution of classical, intermediate and non-classical monocyte subpopulations. Percentages of CD62L+ and CD64+ monocytes were however decreased and increased, respectively, in chronically infected individuals as compared to naïve controls. Infection of monocyte-enriched PBMCs from both seropositive and seronegative individuals with T. gondii led to an increase of CD14+CD16- classical monocytes and a decrease of CD14+CD16+ double positive monocytes. Remarkably, after in vitro parasite infection, expression of the chemokine receptor CCR2 was severely impaired in monocytes from both, individuals with chronic toxoplasmosis and seronegative controls. In contrast, only monocytes from chronically infected humans but not those from controls dose-dependently up-regulated HLA-DR, DP, DQ expression following in vitro infection. Furthermore, monocyte-enriched PBMCs from seropositive individuals up-regulated IL-12 mRNA more vigorously after in vitro infection than cells from naïve controls. Collectively, our results establish that infection of humans with T. gondii exerts long-term effects on the phenotype and responsiveness of blood monocytes. This may have important implications for innate immune responses to T. gondii and unrelated pathogens.
Asunto(s)
Monocitos/inmunología , Toxoplasma/inmunología , Toxoplasmosis/inmunología , Animales , Enfermedad Crónica , Humanos , Inmunidad Innata , Interleucina-12/metabolismo , Selectina L , Receptores de Lipopolisacáridos , Ratones , Monocitos/metabolismo , Receptores CCR2 , Receptores de IgG/inmunología , Toxoplasma/patogenicidadRESUMEN
BACKGROUND: Existential behavioural therapy (EBT) was developed to support informal caregivers of palliative patients in the last stage of life and during bereavement as a manualised group psychotherapy comprising six sessions. We tested the effectiveness of EBT on mental stress and quality of life (QOL). METHODS: Informal caregivers were randomly assigned (1:1) to EBT or a treatment-as-usual control group using computer-generated numbers in blocks of 10. Primary outcomes were assessed with the Brief Symptom Inventory (subscales somatisation, anxiety and depression), the Satisfaction with Life Scale (SWLS), the WHOQOL-BREF and a numeric rating scale for QOL (QOL-NRS, range 0-10). Data were collected at baseline, pre-treatment, post-treatment and follow-ups after 3 and 12 months. Treatment effects were assessed with a multivariate analysis of covariance. RESULTS: Out of 160 relatives, 81 were assigned to EBT and 79 to the control group. Participants were 54.5 ± 13.2 years old; 69.9% were female. The multivariate model was significant for the pre-/post-comparison (p=0.005) and the pre-/12-month comparison (p=0.05) but not for the pre-/3-month comparison. Medium to large effects on anxiety and QOL (SWLS, WHOQOL-BREF, QOL-NRS) were found at post-treatment; medium effects on depression and QOL (QOL-NRS) emerged in the 12-month follow-up. No adverse effects of the intervention were observed. CONCLUSION: Existential behavioural therapy appears to exert beneficial effects on distress and QOL of informal caregivers of palliative patients. Further longitudinal evidence is needed to confirm these findings.
Asunto(s)
Terapia Conductista/métodos , Cuidadores/psicología , Existencialismo/psicología , Neoplasias/enfermería , Cuidados Paliativos/psicología , Psicoterapia de Grupo/métodos , Estrés Psicológico/prevención & control , Adulto , Anciano , Ansiedad/prevención & control , Ansiedad/terapia , Aflicción , Depresión/prevención & control , Depresión/terapia , Femenino , Humanos , Masculino , Persona de Mediana Edad , Atención Plena/métodos , Calidad de Vida , Estrés Psicológico/terapia , Resultado del TratamientoRESUMEN
The effect of liquid compressibility on the dynamics of a single, spherical cavitating bubble is studied. While it is known that compressibility damps the amplitude of bubble rebounds, the extent to which this effect is accurately captured by weakly compressible versions of the Rayleigh-Plesset equation is unclear. To clarify this issue, partial differential equations governing conservation of mass, momentum, and energy are numerically solved both inside the bubble and in the surrounding compressible liquid. Radiated pressure waves originating at the unsteady bubble interface are directly captured. Results obtained with Rayleigh-Plesset type equations accounting for compressibility effects, proposed by Keller and Miksis [J. Acoust. Soc. Am. 68, 628-633 (1980)], Gilmore, and Tomita and Shima [Bull. JSME 20, 1453-1460 (1977)], are compared with those resulting from the full model. For strong collapses, the solution of the latter reveals that an important part of the energy concentrated during the collapse is used to generate an outgoing pressure wave. For the examples considered in this research, peak pressures are larger than those predicted by Rayleigh-Plesset type equations, whereas the amplitudes of the rebounds are smaller.
Asunto(s)
Microburbujas , Modelos Teóricos , Ultrasonido , Simulación por Computador , Análisis Numérico Asistido por Computador , Oscilometría , Presión , Propiedades de Superficie , TemperaturaRESUMEN
We determined the CCR5 chemokine receptor and cytochrome P450 aromatase (P450arom) transcript copies number in swim-up sperm isolated from fertile and infertile men. The ejaculates were purified by centrifugation through discontinuous Percoll density gradient and swim-up techniques. RNA was isolated from sperm, treated with DNase I and reverse-transcribed into cDNA. Quantitative analysis of CCR5 and P450arom cDNA were performed by real-time quantitative (RQ-PCR) SYBR Green I analysis. There was a higher content of CCR5 and P450arom transcripts copy number in swim-up sperm of fertile than from infertile donors. The decrease in CCR5 and P450arom transcripts in swim-up sperm may be associated with male infertility.
Asunto(s)
Aromatasa/genética , Fertilidad/genética , Infertilidad Masculina/genética , ARN Mensajero/análisis , Receptores CCR5/genética , Espermatozoides/metabolismo , Humanos , Masculino , Reacción en Cadena de la Polimerasa , ARN Mensajero/genéticaRESUMEN
The present study was carried out to investigate the relationship between sperm subpopulation kinetics on in vitro fertilization rate. The ability of human sperm to achieve fertilization oocytes was investigated in relation to particular motility parameters obtained on a computer aided sperm analysis system base. Analysis covers velocity straight linear (VSL), cross beat frequency (CBF), lateral head displacement (LHD) and homogeneity of progressive motility velocity (HPMV) of fresh semen and semen after density gradient selection. Investigation was based on sperm samples from 82 infertile couples undergoing IVF. Two subpopulations were extracted from each sample using the clustering method with respect to VSL parameter: a slow and rapid one. Comparison of obtained results before and after selection shows no significant change of subpopulations percentage. However, this method of selection strongly influences motility parameters of both subpopulations. There was found a positive correlation for VSL, LHD and HPMV and a negative correlation for CBF parameters found in slow fraction of fresh semen and percentage of fertilized oocytes. On the other hand, rapid subpopulation parameters for fresh semen and parameters found for both subpopulations in semen after selection did not correlate with one. This means that information of slow sperm subpopulation kinetics carries important prognostic value of IVF success. Since the current prognosis factors ignore motility parameters of slow sperms, our results show the importance of such an analysis.
Asunto(s)
Fertilización In Vitro , Índice de Embarazo , Motilidad Espermática/fisiología , Interacciones Espermatozoide-Óvulo/fisiología , Espermatozoides/citología , Adulto , Femenino , Humanos , Cinética , Masculino , Embarazo , Espermatozoides/clasificación , Espermatozoides/fisiologíaRESUMEN
A proportion of fertilized oocytes during classical in vitro fertilization (IVF) procedure was analysed depending on the following factors: number of mature oocytes, seminological criteria such as sperm morphology in raw semen and after its selection in a density gradient (six structural defects of a male gamete were taken into consideration), sperm concentration, motility parameters according to World Health Organization criteria and the functional tests: hypo-osmotic swelling assay and acrosomal reaction induced by calcium ionophore. Evaluation of DNA content in sperm by image cytometry and determination of malonyldialdehydes in seminal plasma were also performed. Seventy-nine semen samples from patients undergoing IVF were assessed. Apart from significant correlations obtained for selected semen parameters and proportion of fertilized eggs, logistic regression analysis showed that the best predictive factors for oocyte fertilization were normal morphology of sperm before and after gradient selection, grade B and C of sperm movement in raw semen, and DNA content after density gradient centrifugation, which all accounted for 76.7% of fertilization predictive value.
Asunto(s)
Fertilización In Vitro , Oocitos/fisiología , Interacciones Espermatozoide-Óvulo , Espermatozoides/fisiología , Humanos , Peroxidación de Lípido , MasculinoAsunto(s)
Translocador 1 del Nucleótido Adenina/genética , Bovinos/genética , Mapeo Cromosómico/veterinaria , Células Híbridas/efectos de la radiación , Hibridación Fluorescente in Situ/veterinaria , Animales , Secuencia de Bases , Cromosomas Artificiales Bacterianos , Cartilla de ADN , Datos de Secuencia MolecularRESUMEN
Patients with gamma heavy chain disease (gamma-HCD) generally produce incomplete immunoglobulin (Ig) gamma-heavy chains (gamma-HCD protein) which cannot associate with light chains (IgL). In most patients Bence Jones proteins (BJP) are not observed. However, in the 61-year-old patient WIN we found gamma l-HCD proteins and lambda BJP in serum and urine. WIN gamma l-HCD protein does not carry the Ig Fd region, has a molecular weight of 33.5 kDa, and the seven N-terminal amino acid residues are not translated from any of the known immunoglobulin heavy chain (IgH) gene sequences. These residues are followed by the C gamma l-hinge region. In DNA from peripheral blood lymphocytes of patient WIN we found bands representing dominant rearrangements in one of the two alleles of the IgH, Ig kappa and Ig lambda locus. Taken together, the data from protein and DNA analysis strongly suggest, albeit do not formally prove, that one dominant B-cell clone which carries a rearranged and a non-rearranged allele of each Ig locus produces gamma-HCD protein and lambda BJP. The productive lambda-gene rearrangement in this clone thus has not been preceded by abortive rearrangements in both kappa-locus alleles. Lymphocytes with an unusual sequence of IgL-chain gene activation seem to be involved in the case of gamma-HCD described here.
Asunto(s)
Reordenamiento Génico de Cadena Ligera de Linfocito B , Enfermedad de las Cadenas Pesadas/inmunología , Cadenas Ligeras de Inmunoglobulina/genética , Cadenas gamma de Inmunoglobulina/genética , Secuencia de Aminoácidos , Southern Blotting , Cromatografía en Gel , Mapeo Cromosómico , ADN/análisis , Humanos , Regiones Constantes de Inmunoglobulina/genética , Cadenas Pesadas de Inmunoglobulina/genética , Masculino , Persona de Mediana Edad , Datos de Secuencia MolecularRESUMEN
We describe a 61-year-old patient suffering from gamma-1-heavy-chain disease (gamma 1-HCD) associated with Bence-Jones-lambda proteinemia and proteinuria. The analysis of the patients gamma 1-HCD protein (WIN) shows a deletion of the complete Fd fragment. The N-terminal seven amino-acid residue does not resemble any of the known immunoglobulin-heavy-chain variable regions. Unexpectedly, in PBL-DNA and in DNA from EBV-immortalized cells we found in addition to the expected predominantly rearranged Ig-lambda-light-chain gene a predominant rearrangement of an Ig-kappa gene. These findings show that the gamma-1-heavy-chain disease of the patient involves a defective regulation of Ig-light-chain-gene activation as well.
Asunto(s)
Proteína de Bence Jones/análisis , Enfermedad de las Cadenas Pesadas/sangre , Secuencia de Aminoácidos , Proteína de Bence Jones/orina , Reordenamiento Génico , Enfermedad de las Cadenas Pesadas/orina , Humanos , Inmunoelectroforesis , Cadenas gamma de Inmunoglobulina/química , Cadenas kappa de Inmunoglobulina/genética , Cadenas lambda de Inmunoglobulina/genética , Masculino , Persona de Mediana Edad , Datos de Secuencia MolecularRESUMEN
Immunoglobulin-(Ig-) and T-cell-receptor-(TcR-)gene rearrangements were investigated in peripheral blood lymphocytes (PBL) of patients with various autoimmune disorders. In patients with SLE there was no predominant Ig- or TcR-gene rearrangement. This was also true in patients with a long disease duration and with excessive hypergammaglobulinemia. These results lead us to suggest that B cells are activated polyclonally in these patients. In those cases, where predominantly rearranged Ig- or TcR-genes were found, the autoimmune disorder was associated with a low-grade non-Hodgkin lymphoma (NHL). This coherence of B-cell malignancy and autoimmunity was only found in patients with cryoglobulinemia (KG), cold agglutinin disease (KA), and hemolytic anemia (AIHA).
Asunto(s)
Enfermedades Autoinmunes/genética , Reordenamiento Génico de Linfocito T , Reordenamiento Génico , Genes de Inmunoglobulinas , Receptores de Antígenos de Linfocitos T/genética , Adulto , Agammaglobulinemia/genética , Agammaglobulinemia/inmunología , Anciano , Anemia Hemolítica Autoinmune/complicaciones , Anemia Hemolítica Autoinmune/genética , Anemia Hemolítica Autoinmune/inmunología , Enfermedades Autoinmunes/complicaciones , Enfermedades Autoinmunes/inmunología , Linfocitos B/inmunología , Crioglobulinemia/complicaciones , Crioglobulinemia/genética , Crioglobulinemia/inmunología , Femenino , Humanos , Lupus Eritematoso Sistémico/genética , Lupus Eritematoso Sistémico/inmunología , Linfocitos/inmunología , Linfoma no Hodgkin/complicaciones , Masculino , Persona de Mediana Edad , Esclerodermia Sistémica/genética , Esclerodermia Sistémica/inmunología , Síndrome de Sjögren/genética , Síndrome de Sjögren/inmunologíaRESUMEN
In a 64-year-old patient with typical common variable immunodeficiency (CVID) DNA prepared from peripheral blood lymphocytes (PBL) showed a prominent immunoglobulin heavy chain (IgH) gene rearrangement; the immunoglobulin light chain (IgL) genes were found to be in the germline configuration. In contrast, a 13-year-old girl with a hitherto unidentified immunodeficiency showed polyclonal IgH gene rearrangements and a dominant IgL gene rearrangement. In both cases neither monoclonal B-lymphocytes nor monoclonal immunoglobulins were detectable. Our explanation for this unusual observation is that V-gene use in a given B-cell is not entirely random. This may be the consequence of a maturation arrest of B-cells.
Asunto(s)
Reordenamiento Génico de Cadena Pesada de Linfocito B , Reordenamiento Génico de Cadena Ligera de Linfocito B , Cadenas Pesadas de Inmunoglobulina/genética , Cadenas Ligeras de Inmunoglobulina/genética , Síndromes de Inmunodeficiencia/genética , Adolescente , Linfocitos B/inmunología , Femenino , Humanos , Persona de Mediana EdadRESUMEN
The majority of common variable immunodeficiencies (CVID) is caused by intrinsic B cell defects which impede distinct stages of B cell differentiation. B cell differentiation is accompanied by the rearrangement of immunoglobulin (Ig) genes. The first step in the rearrangement process is the assembly of IgH genes, and subsequently, IgL genes are rearranged. During B cell maturation, Ig genes are demethylated in a stepwise, locus-specific manner. Here, we examined the Ig gene rearrangements of four patients with classical CVID and of one child suffering from an unusual immunodeficiency associated with CD5+ B cell lymphocytosis. In one of the four adult patients with CVID, we observed a predominant type of VHDJH-gene rearrangement. In the child, different polyclonal VHDJH-gene rearrangements were found together with a predominant type of kappa light chain gene rearrangement. The rearranged kappa chain genes were methylated (as in the pre-B cell stage). These findings together with the cell phenotype analysis and the clinical course of the disease in the child suggests that in some patients with primary immunodeficiency a maturation arrest may occur in B cells leading to a predominant Ig V gene rearrangement.
Asunto(s)
Reordenamiento Génico , Genes de Inmunoglobulinas , Región Variable de Inmunoglobulina/genética , Síndromes de Inmunodeficiencia/genética , Adolescente , Adulto , Linfocitos B/patología , Southern Blotting , Diferenciación Celular , ADN/análisis , Femenino , Humanos , Síndromes de Inmunodeficiencia/inmunología , Masculino , Metilación , Persona de Mediana EdadRESUMEN
The case of a 57-year-old patient with a plasmablastic lymphoma is described. The primary diagnosis was an ordinary plasmacytoma, type IgG kappa. The final course of the disease involved almost all of the organs. The clinical aspect was determined by multiple skin infiltrations. The present report has in common with other published cases the fact that multiple secondary skin involvement of immunoglobulin-secreting tumors, regardless of their origin, have a very poor prognosis. In comparison, primary multiple cutaneous immunoglobulin-secreting tumors have a better prognosis.
Asunto(s)
Inmunoglobulina G , Cadenas kappa de Inmunoglobulina , Mieloma Múltiple/metabolismo , Neoplasias Cutáneas/metabolismo , Anciano , Humanos , Masculino , PronósticoRESUMEN
Human immunoglobulin (Ig) genes are rearranged in an ordered sequence of events during B-cell differentiation: starting at the IgH locus, a productive VHDJH rearrangement leads to the expression of mu chains. Light-chain gene rearrangements have been found in pre-B cells which express mu chains. In these cells rearrangements of Ig kappa light-chain genes precede that of lambda genes. In an IgD/lambda-producing plasmocytoma, however, we found an apparent exception to this rule: the kappa genes were not rearranged. Together with the observation that roughly 90% of human IgD plasmocytomas produce lambda light-chain proteins, the finding reported here leads us to suggest that lambda light-chain genes are rearranged preferentially in IgD-producing plasma cells. Ig gene rearrangement, isotype switch, and the phenomenon of isotypic and allelic exclusion are discussed with special reference to our findings.