Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 36
Filtrar
Mais filtros











Intervalo de ano de publicação
1.
Pediatr. aten. prim ; 22(88): 423-433, oct.-dic. 2020. tab, graf
Artigo em Espanhol | IBECS | ID: ibc-196406

RESUMO

Hasta ahora, los datos recogidos en los casos de procesos clínicos provocados por el coronavirus SARS-CoV-2 (COVID-19) en niños sugieren que son cuadros leves en comparación con las infecciones en pacientes adultos; no obstante, se ha informado de casos graves, como el síndrome inflamatorio multisistémico (SIM), que precisa de valoración y actuación de emergencia. En el contexto de la consulta del pediatra de Atención Primaria y coincidiendo con el inicio del curso escolar, en una época en la que habitualmente aumenta la incidencia de procesos como la gripe, infección por el virus respiratorio sincitial (VRS) y otros cuadros respiratorios, es habitual la demanda por síntomas que pueden hacer sospechar cualquiera de estas infecciones. En este sentido, es importante llegar a un diagnóstico que permita el manejo más adecuado del paciente. Epidemiológicamente, de manera que se pueda disminuir la transmisión comunitaria tomando las medidas adecuadas y clínicamente para así poder ponderar el nivel de gravedad y poner en marcha las actuaciones más adecuadas. Dado que no existen escalas válidas que ofrezcan un puntaje para valorar cuál es la actuación más adecuada ante la sospecha de una infección COVID-19, planteamos los beneficios de un algoritmo de decisión clínica que tiene en cuenta las connotaciones epidemiológicas, basado en la gravedad clínica, para ofrecer la atención clínica más adecuada a los pacientes


So far, the data collected in the cases of clinical processes caused by the SARS-CoV-2 (COVID-19) coronavirus in children suggest that they are mild compared to infections in adult patients; However, serious cases such as multisystemic inflammatory syndrome (SIM) have been reported, which requires assessment and emergency action. In the context of the of the Primary Care pediatrician consultation and coinciding with the beginning of the school year, at a time when the incidence of influenza, RSV infection and other respiratory conditions usually increases, consultations for symptoms that can lead to suspect these infections. Therefore, it is important to reach a diagnosis that allows the most appropriate management of the patient and decreasing the community transmission by taking pertinent measures. Given that there are no valid scales that offer a score to assess which is the most appropriate action in the event of a suspected COVID-19 infection, we propose the benefits of a clinical decision algorithm that takes into account epidemiological connotations, based on clinical severity to offer the most appropriate clinical care to patients


Assuntos
Humanos , Recém-Nascido , Lactente , Pré-Escolar , Criança , Adolescente , Atenção Primária à Saúde , Tomada de Decisão Clínica , Infecções por Coronavirus/terapia , Infecções por Coronavirus/diagnóstico , Pneumonia Viral/terapia , Pneumonia Viral/diagnóstico , Pandemias , Algoritmos
3.
Eur J Pharm Biopharm ; 96: 152-61, 2015 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-26209123

RESUMO

Coated pellets and mini-tablets were prepared containing a new broad spectrum antibacterial agent: CIN-102, a well-defined, synergistic blend of trans-cinnamaldehyde, trans-2-methoxycinnamaldehyde, cinnamyl acetate, linalool, ß-caryophyllene, cineol and benzyl benzoate. The aim was to provide a new treatment method for colitis, especially for Inflammatory Bowel Disease (IBD) patients. Since the simple oral gavage of CIN-102 was not able to reduce the pathogenic bacteria involved in colitis (rat model), the drug was incorporated into multiparticulates. The idea was to minimize undesired drug release in the upper gastrointestinal tract and to control CIN-102 release in the colon, in order to optimize the resulting antibiotic concentration at the site of action. A particular challenge was the fact that CIN-102 is a volatile hydrophobic liquid. Pellet cores were prepared by extrusion-spheronization and coated with polymer blends, which are sensitive to colonic bacterial enzymes. Mini-tablets were prepared by direct compression. The release of the main compound of CIN-102 (cinnamaldehyde, 86.7% w/w) was monitored in vitro. Optimized coated pellets and mini-tablets were also tested in vivo: in seven-week-old, male mice suffering from dextran sodium sulfate induced colitis. Importantly, both types of multiparticulates were able: (i) to significantly reduce the number of luminal and mucosal enterobacteria in the mice (the levels of which are increased in the disease state), and (ii) to improve the clinical course of the intestinal inflammation (decrease in the percentages of mice with bloody stools and diarrhea). Thus, the proposed coated pellets and matrix mini-tablets allowing for controlled CIN-102 release show a promising potential for new treatment methods of colitis.


Assuntos
Antibacterianos/administração & dosagem , Benzoatos/administração & dosagem , Cinamatos/administração & dosagem , Colite/tratamento farmacológico , Sistemas de Liberação de Medicamentos/métodos , Terpenos/administração & dosagem , Administração Oral , Animais , Antibacterianos/química , Antibacterianos/uso terapêutico , Benzoatos/química , Benzoatos/uso terapêutico , Varredura Diferencial de Calorimetria , Cinamatos/química , Cinamatos/uso terapêutico , Colite/microbiologia , Colo/efeitos dos fármacos , Colo/microbiologia , Combinação de Medicamentos , Composição de Medicamentos , Liberação Controlada de Fármacos , Masculino , Camundongos Endogâmicos C57BL , Solubilidade , Propriedades de Superfície , Comprimidos , Terpenos/química , Terpenos/uso terapêutico
4.
Eur J Clin Microbiol Infect Dis ; 27(6): 451-9, 2008 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-18246375

RESUMO

Recently, the incidence of human mycobacterial infections due to species other than M. tuberculosis has increased worldwide. Since disease control depends on appropriate antimicrobial therapy, the precise identification of these species of clinical importance has become a major public health concern. Identification of mycobacteria has been hampered because of the lack of specific, rapid, and inexpensive methods. Therefore, we aimed at designing and validating a bacterial lysate-based polymerase chain reaction identification scheme. This scheme can classify clinical isolates into: (1) the genus Mycobacterium, (2) the M. tuberculosis complex, (3) the nontuberculous mycobacteria, and (4) the species M. avium, M. intracellulare, M. abscessus, M. chelonae, M. fortuitum and M. bovis of clinical importance, and M. gordonae, the most commonly encountered nonpathogenic species in clinical laboratories. By using M. fortuitum and M. avium lysates as models, the method sensitivity was determined to be 372 pg of DNA. In a blind parallel comparison between our approach and conventional biochemical tests, both assays correctly categorized 75 patient's mycobacterial isolates. However, our approach only required 4-9 h for categorization compared with at least 15 days by conventional tests. Furthermore, our methodology could also detect M. fortuitum and M. avium from liquid cultures, after only 2 and 6 days, respectively, of incubation. Our new identification scheme is therefore sensitive, specific, rapid, and economic. Additionally, it can help to provide proper treatment to patients, to control these diseases, and to improve our knowledge of the epidemiology of mycobacteriosis, all urgently needed, particularly in developing countries.


Assuntos
Bactérias/genética , Extratos Celulares/genética , Mycobacterium/patogenicidade , Sensibilidade e Especificidade , Técnicas de Tipagem Bacteriana , Humanos , Mycobacterium/classificação , Mycobacterium/genética , Mycobacterium/isolamento & purificação , Infecções por Mycobacterium/microbiologia , Infecções por Mycobacterium/fisiopatologia , Mycobacterium tuberculosis/genética , Mycobacterium tuberculosis/isolamento & purificação , Reação em Cadeia da Polimerase/métodos , Especificidade da Espécie
5.
Ortod. esp. (Ed. impr.) ; 42(3): 117-124, sept. 2002.
Artigo em Es | IBECS | ID: ibc-23528

RESUMO

Presentamos un estudio cefalométrico sobre una muestra seleccionada de 22 pacientes sometidos a cirugía de los maxilares para la corrección de la Clase III esquelética. Todos ellos estaban fuera de las edades activas de crecimiento facial. De ellos, en seis se llevó a cabo avance unitario del maxilar superior, en otros seis retrusión aislada de la mandíbula y en los restantes 10 cirugía combinada. Se utilizó un análisis cefalométrico que permite el estudio de las relaciones sagitales y verticales de la cara, la estructura nasal y el grosor de los tejidos blandos del perfil facial. Las mediciones fueron efectuadas a partir de telerradiografías laterales de cráneo tomadas antes y después de la intervención quirúrgica. Los resultados así obtenidos se procesaron para su estudio estadístico, en el que realizamos descripción básica, test "t", correlaciones y comparaciones entre los tres grupos de cirugía. Las conclusiones que apuntamos reflejan la corrección conseguida tras el tratamiento a nivel esquelético y del tejido blando facial, así como el comportamiento diferencial de estas estructuras según el tipo de cirugía realizada. (AU)


Assuntos
Adolescente , Adulto , Feminino , Masculino , Humanos , Má Oclusão Classe III de Angle/cirurgia , Má Oclusão Classe III de Angle/diagnóstico , Cefalometria/métodos , Maxila/cirurgia , Anormalidades Maxilomandibulares/cirurgia , Anormalidades Maxilomandibulares/diagnóstico , Maxila/cirurgia
6.
NMR Biomed ; 14(3): 210-6, 2001 May.
Artigo em Inglês | MEDLINE | ID: mdl-11357187

RESUMO

High resolution magnetic resonance imaging (MRI) was applied to quantify alterations in thymus and adrenal volumes, as well as body fat in genetically engineered corticotropin-releasing factor (CRF)-overexpressing mice. When compared to the organs in age-matched wild-type animals, the adrenals in CRF-overexpressing male mice were significantly enlarged and the thymus volume in females was significantly smaller. The fat content was significantly larger in CRF-overexpressing mice. The anatomical alterations observed in the MRI studies were in perfect line with post-mortem data (weights of organs). Furthermore, the observed interstrain differences are in agreement with recently published data on (i) the effect of continuous, intraventricular infusion of CRF in rats and (ii) the presence of atrophic adrenals in CRF-knockout mice. The present studies demonstrate that MRI can provide reliable measures of relatively small structures such as the adrenal glands and the thymus in mice. This makes MRI an attractive, non-terminal tool to monitor in laboratory animals, including transgenic mice, the consequence of continuous stress on relevant organs.


Assuntos
Tecido Adiposo/anatomia & histologia , Glândulas Suprarrenais/anatomia & histologia , Hormônio Liberador da Corticotropina/genética , Hormônio Liberador da Corticotropina/fisiologia , Imageamento por Ressonância Magnética , Timo/anatomia & histologia , Córtex Suprarrenal/patologia , Medula Suprarrenal/anatomia & histologia , Animais , Feminino , Expressão Gênica , Hipertrofia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos
7.
NMR Biomed ; 14(3): 217-22, 2001 May.
Artigo em Inglês | MEDLINE | ID: mdl-11357188

RESUMO

In order to keep subscribers up-to-date with the latest developments in their field, John Wiley & Sons are providing a current awareness service in each issue of the journal. The bibliography contains newly published material in the field of NMR in biomedicine. Each bibliography is divided into 9 sections: 1 Books, Reviews ' Symposia; 2 General; 3 Technology; 4 Brain and Nerves; 5 Neuropathology; 6 Cancer; 7 Cardiac, Vascular and Respiratory Systems; 8 Liver, Kidney and Other Organs; 9 Muscle and Orthopaedic. Within each section, articles are listed in alphabetical order with respect to author. If, in the preceding period, no publications are located relevant to any one of these headings, that section will be omitted.


Assuntos
Espectroscopia de Ressonância Magnética
8.
Lab Anim ; 34(4): 351-61, 2000 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-11072855

RESUMO

Blood samples were taken from the retrobulbar venous plexus or the sublingual vein of male HamIbm:Wist rats to compare clinical pathology parameters between the two sampling techniques. By analogy with a pharmacokinetic study, blood was sampled six times during one day from unfasted animals. After 3 weeks of recovery, blood was taken from fasted animals on a single occasion. In addition, prolactin and corticosterone levels were determined to compare stress-related effects between the two sampling methods. Body weight development and food consumption were similar after single as well as after repeated blood sampling for the two blood sampling techniques. Haemotological evaluation showed a gradual decrease in erythrocyte count, haemoglobin concentration and haematocrit after repeated blood sampling. Repeated withdrawal of blood samples over 24 h corresponding to approximately 22% of the total blood volume resulted in a decrease in red blood cell parameters by up to 30%. The withdrawal of approximately 10% of the total blood volume was associated with a decrease in these parameters by up to 10% and should not be exceeded for animal welfare reasons and to allow a reliable evaluation of data in a study. Repeated blood sampling was associated with an initial decrease in the number of white blood cells, mainly due to a reduction in lymphocytes; white blood cell counts were slightly increased one day after. The decrease in lymphocytes and the increase in neutrophils after repeated sampling were generally slightly more pronounced in the blood from the retrobulbar plexus than from the sublingual vein. Comparison of serum clinical chemistry data showed significantly higher activities of creatine kinase and aspartate aminotransferase in samples from the retrobulbar plexus. These findings suggest a higher degree of tissue damage with blood sampling from the retrobulbar plexus than from the sublingual vein. Despite a large inter-individual variability, higher mean values of prolactin on each occasion and corticosterone after a single sample in fasted animals indicate a higher stress associated with blood sampling from the retrobulbar plexus.


Assuntos
Coleta de Amostras Sanguíneas/veterinária , Ratos/sangue , Estresse Fisiológico/veterinária , Amilases/sangue , Animais , Glicemia/análise , Coleta de Amostras Sanguíneas/métodos , Peso Corporal , Corticosterona/sangue , Creatina Quinase/sangue , Ingestão de Alimentos , Ingestão de Energia , Contagem de Eritrócitos/veterinária , Olho/irrigação sanguínea , Contagem de Leucócitos/veterinária , Masculino , Soalho Bucal/irrigação sanguínea , Prolactina/sangue , Estresse Fisiológico/sangue , Veias
9.
Biochem Pharmacol ; 59(3): 267-72, 2000 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-10609555

RESUMO

Cyclosporine A (CsA) is a potent immunosuppressant with the drawback of renal side effects. We reported that CsA markedly decreases calcium-binding protein calbindin-D28k mRNA levels in rat kidneys, and showed that this decrease is associated with its adverse renal effects. The transcription of the calbindin-D28k gene is activated via the vitamin D pathway. In this work, the potential CsA-mediated impairment of the vitamin D pathway was investigated. Wistar rats were treated for 12 days with 50 mg/kg/day CsA or for 20 days with 50 mg/kg/day of the non-immunosuppressant and non-nephrotoxic SDZ PSC 833, which had been previously shown not to affect calbindin-D28k mRNA levels. The expression of the three vitamin D-regulated genes calbindin-D28k, 1,25-dihydroxyvitamin D3-24-hydroxylase (24-OHase), and vitamin D receptor (VDR) were quantified in rat kidney homogenates by real-time reverse transcription-polymerase chain reaction. Plasma parathyroid hormone (PTH) as well as plasma and kidney 1,25 dihydroxyvitamin D3 (calcitriol) levels were monitored in all animals. CsA induced a 85% decrease in calbindin-D28k mRNA levels as well as a 40% and 69% decrease in VDR and 24-OHase mRNA levels, respectively. Plasma and kidney 1,25 dihydroxyvitamin D3 as well as plasma PTH levels were increased by CsA, but not by SDZ PSC 833. The treatment with SDZ PSC 833 did not affect calbindin-D28k or VDR expression, but did cause a 73% decrease in 24-OHase mRNA levels. Taken together, these results indicate an association between CsA-mediated down-regulation of rat renal calbindin-D28k mRNA and the decrease in other 1,25 dihydroxyvitamin D3-regulated genes, suggesting an impairment of the vitamin D pathway by CsA which may be related to its adverse renal side effects and its immunosuppressive activity.


Assuntos
Ciclosporina/farmacologia , Imunossupressores/farmacologia , Insuficiência Renal/metabolismo , Vitamina D/metabolismo , Animais , Ciclosporina/efeitos adversos , Ciclosporinas/efeitos adversos , Ciclosporinas/farmacologia , Expressão Gênica/efeitos dos fármacos , Imunossupressores/efeitos adversos , Masculino , Ratos , Ratos Wistar , Receptores de Calcitriol/genética , Receptores de Calcitriol/metabolismo , Insuficiência Renal/induzido quimicamente , Insuficiência Renal/genética
10.
Reprod Toxicol ; 12(1): 39-47, 1998.
Artigo em Inglês | MEDLINE | ID: mdl-9431571

RESUMO

In this study, the suitability of several methods for the assessment of testicular damage, including histopathology, flow cytometry (FCM), testicular sperm head counts, and secretion of androgen binding protein (ABP), has been evaluated. Testicular toxicity after acute exposure of adult rats to different doses of the known toxicant 1,3-dinitrobenzene (DNB) was analyzed. The effects showed dose dependence, in spite of the large variability within each dose group. Histopathology and FCM showed germ cell depletion, particularly of round spermatids; testicular sperm head counts were reduced and ABP production was increased. All evaluated methods showed similar sensitivities. The increased testicular ABP levels support the theory that the Sertoli cell is the likely target of DNB induced testicular toxicity, producing subsequent germ cell depletion. The presented results show the suitability of FCM for the analysis of testicular damage and also support the usefulness of including a metabolic marker for Sertoli cell function.


Assuntos
Doenças Testiculares/induzido quimicamente , Toxicologia/métodos , Proteína de Ligação a Androgênios/análise , Animais , Peso Corporal/efeitos dos fármacos , Dinitrobenzenos/toxicidade , Citometria de Fluxo , Masculino , Tamanho do Órgão/efeitos dos fármacos , Ratos , Ratos Wistar , Células de Sertoli/efeitos dos fármacos , Contagem de Espermatozoides/efeitos dos fármacos , Cabeça do Espermatozoide/efeitos dos fármacos , Doenças Testiculares/metabolismo , Doenças Testiculares/patologia , Testículo/anatomia & histologia , Testículo/efeitos dos fármacos
11.
Arch Toxicol ; 72(12): 791-7, 1998 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-9950076

RESUMO

The aim of this study was to compare results obtained by flow cytometry (FCM) with those obtained from testicular histopathology with regard to testicular damage following acute exposure of adult rats to the known testicular toxicant, methoxyacetic acid (MAA). Special emphasis was given to defining the sensitivity of three-parameter FCM compared with testicular histopathology. Furthermore, the effect on the male reproductive system of a single oral dose of MAA was evaluated with traditional methods, e.g. testicular sperm head counts, and organ weights. Adult, male Han/Wistar rats were randomly assigned to four groups of ten animals to be treated with a single dose of 0, 65, 325 and 650 mg MAA/kg body wt. (p.o., gavage). The animals were killed 2 days after treatment, and testicular and epididymal weights were recorded. One testis and the corresponding epididymis were used for histopathology. The other testis was used partly to determine sonication-resistant, testicular sperm-head counts (SHC), and partly for enzymatic digestion followed by FCM. The results obtained in this study are in agreement with the literature, and show that, in the adult male rat, 2 days after administering a single oral dose of MAA, specific depletion of spermatocytes is evident. Detectable testicular effects were produced by the high (650 mg/kg body wt.) and mid (325 mg/kg body wt.) doses, whilst the low dose (65 mg/kg body wt.) did not produce any noticeable effect. There was a strong correlation between results obtained by FCM and those obtained by testicular histopathology, and no difference in sensitivity between the two methods was observed. In summary, three-parameter FCM represents a sensitive and reliable method for the detection of testicular injury in the rat. It requires only small amounts of tissue, and the sensitivity was shown to be similar to that of histopathology. Moreover, FCM has the advantages of being quick and objective, which permits large numbers of cells to be analysed. The potential use of this method as a fast screening tool for testicular toxicity in routine toxicology studies should be considered.


Assuntos
Acetatos/toxicidade , Citometria de Fluxo , Testículo/efeitos dos fármacos , Testes de Toxicidade/métodos , Administração Oral , Animais , Epididimo/efeitos dos fármacos , Epididimo/patologia , Masculino , Tamanho do Órgão/efeitos dos fármacos , Ratos , Ratos Wistar , Sensibilidade e Especificidade , Cabeça do Espermatozoide/efeitos dos fármacos , Testículo/patologia
13.
Environ Mol Mutagen ; 32(4): 387-96, 1998.
Artigo em Inglês | MEDLINE | ID: mdl-9882014

RESUMO

Measurement of the frequency of micronuclei induced in cells by ionizing radiation or by chemical treatment is widely used to analyze cytogenetic damage. The microscopic scoring of micronuclei is a tedious and time-consuming procedure. Therefore, attempts have been made to automate micronuclei scoring by means of image analysis or flow cytometry. A new procedure for the flow cytometric analysis of chemically induced micronuclei in V79 Chinese hamster cells has been established in our laboratory. Debris was separated from micronuclei by means of a new gating procedure using area and width fluorescence of the stained suspension of micronuclei and nuclei. In order to test the sensitivity and specificity of this improved method of flow cytometric analysis, five well-known mutagenic compounds were tested. With the new technique, the frequency of micronuclei measured and analyzed corresponded well with results obtained by conventional microscopy. In addition, a large series of negative compounds, and weak, middle, and strong micronuclei inducers, were tested in order to establish criteria for discrimination between genotoxic and nongenotoxic compounds by flow cytometry. This new procedure for flow cytometric detection of micronuclei represents a quick, reliable, and relatively simple method for in vitro micronucleus testing.


Assuntos
Citometria de Fluxo/métodos , Testes para Micronúcleos/métodos , Mutagênicos/toxicidade , Animais , Bleomicina/toxicidade , Linhagem Celular , Cricetinae , Cricetulus , Ciclofosfamida/toxicidade , Metanossulfonato de Etila/toxicidade , Estudos de Avaliação como Assunto , Vimblastina/toxicidade , Vincristina/toxicidade
14.
J Clin Microbiol ; 35(12): 3048-53, 1997 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-9399492

RESUMO

While probing the vicinity of ompC, a copy of the IS200 insertion element was found between the gyrA and rcsC genes of Salmonella typhi, the causal agent of typhoid fever. This distinctive feature was conserved throughout 63 S. typhi isolates of different geographical origins and was absent from 46 other Salmonella serotypes, including those most associated with human infections, as well as from 19 other enteric bacteria. Furthermore, the location of this IS200 copy corresponds to a constant band, present throughout the 14 PstI S. typhi IS200 fingerprints, encompassing several Vi phage types. Interestingly, an apparently unrelated serotype not frequently associated with human disease, Salmonella weltevreden, contained an IS200 copy at the same position, although it was accompanied by an additional segment of cryptic DNA. On the basis of these findings, PCR assays were designed for molecular typing of S. typhi, and these are potentially useful in studying the epidemiology of typhoid fever.


Assuntos
Elementos de DNA Transponíveis , Genes Bacterianos , Complexos Multienzimáticos , Fosfoproteínas Fosfatases , Proteínas Quinases , Salmonella typhi/genética , Proteínas de Bactérias/genética , Sequência de Bases , Mapeamento Cromossômico , Sequência Conservada , Impressões Digitais de DNA , DNA Girase , Primers do DNA/genética , DNA Topoisomerases Tipo II/genética , DNA Bacteriano/genética , DNA Bacteriano/isolamento & purificação , Genoma Bacteriano , Humanos , Epidemiologia Molecular , Dados de Sequência Molecular , Reação em Cadeia da Polimerase , Salmonella typhi/classificação , Salmonella typhi/isolamento & purificação , Sorotipagem , Fatores de Transcrição/genética , Febre Tifoide/epidemiologia , Febre Tifoide/microbiologia
15.
Reprod Toxicol ; 11(4): 521-31, 1997.
Artigo em Inglês | MEDLINE | ID: mdl-9241672

RESUMO

Histopathologic examination of testicular tissue allows testicular impairment to be investigated. As an alternative to histopathology, flow cytometry (FCM) using a triple staining technique that combines DNA-ploidy with mitochondria stainability and vimentin immunostaining has also been utilized to evaluate testicular damage. In this article we evaluate the effects on spermatogenesis after acute exposure of rats to doxorubicin. Testicular cell suspensions of treated and control animals were analyzed by FCM. This allows several cell types to be identified and quantified, giving a control pattern. Deviations from this control pattern are considered as an indication of testicular damage. Doxorubicin produced a depletion of spermatogonia as early as 3 d after treatment. This effect could be followed through the temporal evolution of spermatogenesis. Comparable results were obtained by histopathology. The presented results show that FCM is a suitable and sensitive method for the detection of testicular damage. The advantages of FCM over other techniques include its rapidity and objectivity.


Assuntos
Antibióticos Antineoplásicos/toxicidade , Doxorrubicina/toxicidade , Espermatogênese/efeitos dos fármacos , Animais , Peso Corporal/efeitos dos fármacos , Citometria de Fluxo , Masculino , Tamanho do Órgão/efeitos dos fármacos , Ratos , Ratos Wistar , Contagem de Espermatozoides/efeitos dos fármacos , Testículo/efeitos dos fármacos
16.
Cytometry ; 27(2): 161-8, 1997 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-9012383

RESUMO

Mammalian spermatogenesis is a complex process which is not yet fully understood. In this paper we describe the analysis of rat spermatogenesis by means of 3-parameter flow cytometry. Since the analysis of DNA content only provides sufficient information for the identification of 4 cell populations, additional parameters were combined with propidium iodide (PI) staining. Immunostaining of the intermediate filament vimentin allowed the identification of somatic (vimentin positive) and germ (vimentin negative) cells. Utilizing the combination of DNA and vimentin staining, we have been able to quantitate the somatic cells present in a testicular cell suspension and to analyze somatic and germinal cells separately. Furthermore, the addition of mitochondrial staining with the fluorochrome nonyl acridine orange (NAO) allowed several cell subpopulations within each ploidy group to be distinguished. After 3-color staining and subsequent cell sorting, 11 testicular cell subpopulations could be identified: somatic cells, and 10 subtypes of germinal cells. The method described in this paper represents a valuable tool for the evaluation of spermatogenesis in both normal and perturbed situations.


Assuntos
Citometria de Fluxo/métodos , Espermatogênese , Testículo/citologia , Laranja de Acridina/química , Animais , DNA/análise , Masculino , Ficoeritrina/química , Propídio/química , Coelhos , Ratos , Ratos Wistar , Reprodutibilidade dos Testes , Espermatogênese/fisiologia , Vimentina/metabolismo
17.
Hum Gene Ther ; 7(4): 531-6, 1996 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-8800748

RESUMO

The in vivo gene transfer to mouse hepatocytes of pTG 7101, a plasmid containing the full-length gene encoding human alpha 1-antitrypsin (alpha 1-AT) DNA, has been studied by iv administration of recombinant DNA (100 ng/mouse) encapsulated in large and small liposomes. Our results from immunohistochemical liver sections and cytophotometric analysis of hepatocyte chromophore absorbance indicate that human alpha 1-AT was expressed in liver parenchymal cells from mice treated (48 hr before) with DNA encapsulated in small liposomes, and this effect remained for at least 2 weeks. In contrast, the efficiency was greatly limited when large liposomes were used as a vehicle for gene transfer. Additional experiments were performed to study using an ELISA procedure the presence in mouse plasma of human alpha 1-AT from mice treated with encapsulated plasmid in small liposomes or small empty liposomes plus free DNA. According to the immunohistochemical data, the results indicate that detectable alpha 1-AT can only be observed in plasma from mice treated with encapsulated plasmid in small liposomes.


Assuntos
Técnicas de Transferência de Genes , Fígado/metabolismo , alfa 1-Antitripsina/genética , Animais , Humanos , Lipossomos/metabolismo , Fígado/citologia , Camundongos , alfa 1-Antitripsina/metabolismo
18.
Gene ; 165(1): 1-8, 1995 Nov 07.
Artigo em Inglês | MEDLINE | ID: mdl-7489896

RESUMO

Campylobacter jejuni (Cj) and C. coli (Cc) clinical isolates, obtained from three different sources, were characterized using two Cj DNA probes, CJ01 and CJ02. These probes were selected at random by virtue of their stability in Escherichia coli (Ec). CJ01 hybridized specifically with DNA from Cj reference strains, but not with DNA from Cc, C. lari (Cl) nor C. fetus (Cf) reference strains. Using clinical isolates characterized by genome-genome hybridization and biotype, CJ01 hybridized with DNA derived from all Cj strains. However, DNA from four out of ten Cc strains, from three different sources, also hybridized with CJ01, suggestive of this region being heterogeneous between clinical isolates of both species. The nucleotide sequence analysis of CJ01 reveals two incomplete open reading frames (ORFs) that did not show significant homology with any other known sequences. CJ02 hybridized specifically with DNA from Cj and Cc reference strains, but not with DNA from Cl and Cf reference strains. The specificity and sensitivity were maintained upon hybridization with DNA from 31 clinical isolates. CJ02 has an uninterrupted ORF whose deduced amino-acid sequence showed extensive homology with the central region of the Ec and Salmonella typhimurium (St) RNA polymerase beta subunits (52 and 66% similarity, respectively). The most conserved segments correspond to putative functional domains.


Assuntos
Proteínas de Bactérias/genética , Campylobacter coli/genética , Campylobacter jejuni/genética , Sequência de Aminoácidos , Técnicas de Tipagem Bacteriana , Sequência de Bases , Campylobacter coli/classificação , Campylobacter coli/isolamento & purificação , Campylobacter jejuni/classificação , Campylobacter jejuni/isolamento & purificação , Sondas de DNA , Genes Bacterianos , Dados de Sequência Molecular , Sensibilidade e Especificidade
19.
Immunopharmacol Immunotoxicol ; 17(3): 419-36, 1995 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-8576538

RESUMO

Immunization of C57BL/6 mice with tumor-derived membrane-proteins encapsulated in sized liposomes (0.2 microgram/mouse) and composed by phosphatidylcholine or sphingomyelin, significantly reduced the mean values of spontaneous lung metastasis from both B16 (0.7 +/- 0.5 and 1.2 +/- 0.6, respectively) and 3LL (4.8 +/- 2.5 and 7.2 +/- 4.1, respectively) tumors, with respect to control (HEPES) groups (4.8 +/- 1.1 and 19.0 +/- 4.4, respectively). However, no significant antimetastatic effect was observed using free tumor-derived proteins (2 micrograms/mouse) or liposome vehicle alone. Specific humoral immune response after the vaccination was studied by flow cytometry of tumor cells incubated with a pooled sample from each group of immunized mice and FITC-conjugate antimouse immunoglobulins. The results showed that the highest number of positive tumor cells was identified using sera from immunized mice with sized liposomes encapsulating tumor-derived proteins whereas the immunization with the protein fraction in free form failed to induce this effect. In addition, an increased cytotoxicity towards 3LL and B16 tumor cells can also be observed when tumor cells were incubated with spleen effector cells plus specific immunosera. In conclusion, our results show that antitumor active vaccination, using sized liposomes as adjuvants, induces an antitumor host response and a significant inhibition of tumor progression.


Assuntos
Antígenos de Neoplasias/administração & dosagem , Imunização , Neoplasias Pulmonares/prevenção & controle , Neoplasias Pulmonares/secundário , Animais , Citotoxicidade Celular Dependente de Anticorpos , Antígenos de Superfície/administração & dosagem , Membrana Celular/imunologia , Técnicas In Vitro , Lipossomos , Neoplasias Pulmonares/imunologia , Masculino , Melanoma Experimental/imunologia , Melanoma Experimental/secundário , Melanoma Experimental/terapia , Camundongos , Camundongos Endogâmicos C57BL , Baço/imunologia
20.
Gene ; 156(1): 1-9, 1995 Apr 14.
Artigo em Inglês | MEDLINE | ID: mdl-7537703

RESUMO

The Salmonella typhi (St) ompC gene codes for a major outer membrane protein (OMP) that is highly expressed in both low and high osmolarity. By hybridization studies with the entire gene or with segments thereof, ompC was found to be highly conserved within 11 different Salmonella serotypes, with the exception of S. arizonae. The study included several St isolates from Mexico and Indonesia. Variation was only detected in two (e and f) of the seven regions previously found to vary between St and E. coli ompC. Chimeric OmpC proteins, carrying a rotavirus VP4 capsid protein epitope, are well recognized by a specific monoclonal antibody (mAb) against this epitope, either in OMP preparations (by enzyme-linked immunosorbent assay; ELISA) or intact cells (by ELISA and immunogold-labelling), indicating that regions c and f are oriented towards the cell surface and are probably exposed. As has been shown before for other regulated OMP, this experimental approach could be useful for the presentation of heterologous epitopes in order to gain knowledge about porin topology, for testing the effect of altered porin surface epitopes on bacterial physiology, or else, in the development of multivalent vaccines.


Assuntos
Proteínas da Membrana Bacteriana Externa/genética , Proteínas do Capsídeo , Epitopos/análise , Genes Bacterianos/genética , Polimorfismo de Fragmento de Restrição , Salmonella typhi/genética , Sequência de Aminoácidos , Proteínas da Membrana Bacteriana Externa/química , Proteínas da Membrana Bacteriana Externa/imunologia , Sequência de Bases , Capsídeo/análise , Capsídeo/genética , Sequência Conservada/genética , Epitopos/química , Variação Genética/genética , Genótipo , Dados de Sequência Molecular , Conformação Proteica , Proteínas Recombinantes de Fusão/biossíntese , Proteínas Recombinantes de Fusão/imunologia , Rotavirus/genética , Rotavirus/imunologia , Salmonella/genética
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA