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1.
Transplant Proc ; 47(7): 2097-101, 2015 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-26361652

RESUMO

BACKGROUND: Health workers' awareness and knowledge of transplant medicine can improve people's sensitivity and reduce their degree of opposition to donations. The medical literature contains numerous examples of education programs aimed at university students. This work describes the experience of an education program for students of the second and third year of a nursing degree course. METHODS: From April to September 2013, an education program was set up for 80 university students. It was divided into 3 stages: group self-learning based on prearranged topics, sharing of the results, and participation in the final seminar. The effectiveness was assessed according to a pretest/posttest design. RESULTS: The first questionnaire contained 19 questions, and the second contained 27. The questions were subdivided into specific areas: subjective knowledge, objective knowledge, attitude, awareness, participation in the event, evaluation of the information material handed out, and appreciation of the tools used. There was a significant increase for items relating to knowledge, whereas awareness and attitude (already high at the start of the program) showed no changes. After the program, many students discussed the question of donation with their relatives and friends, and about 70% filled in a donor card. The students expressed a highly positive opinion of the initiative and the tools used. CONCLUSIONS: The initiative proved its validity, improving subjective and objective knowledge to a statistically significant extent and also increasing awareness and attitude. The students' evaluation was extremely positive.


Assuntos
Atitude do Pessoal de Saúde , Educação em Enfermagem/métodos , Conhecimentos, Atitudes e Prática em Saúde , Transplante de Órgãos/educação , Estudantes/psicologia , Doadores de Tecidos , Adulto , Feminino , Humanos , Masculino , Avaliação de Programas e Projetos de Saúde , Inquéritos e Questionários , Obtenção de Tecidos e Órgãos , Adulto Jovem
2.
Transplant Proc ; 45(7): 2584-6, 2013 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-24033996

RESUMO

Between 2004 and 2010 in Piedmont (Italy Northern Region) 1556 brain-death situations were reported, including 113 (7.3%) in migrants as potential organ and tissue donors. The health staff often has to face migrants, who show great cultural differences and language difficulties. The Molinette Hospital Customer Care Service, the Piedmont Regional Tissue and Organ Procurement Coordination Agency (RPC), and the Cross-Cultural Mediators Association (CMA) organized a special course for intercultural mediators, to decrease misunderstandings between the health staff and the migrants' families and to improve professional communication. In 2011, 28 cultural-linguistic mediators representing different groups of migrants in Piemonte took part in a specific course. Over a 5 month period they were informed about emotional and communicative aspects, proper to the moment of death, as well as organ donation as an intercultural field, the professional role of the mediator, the clinical and forensic aspects of brain death and donation, and the psychological aspects of organ donation. The course was organized by cultural-linguistic mediators of the CMA, the staff of the RPC and the teachers at Turin University. The list of the 21 mediators who passed the final exam was given to organ and tissue donation hospital co-ordinators in Piedmont, so that if necessary, they could obtain the cooperation of these qualified people.


Assuntos
Comparação Transcultural , Obtenção de Tecidos e Órgãos , Migrantes , Humanos
3.
Transplant Proc ; 45(7): 2580-3, 2013 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-24033995

RESUMO

FOREWORD: Law no. 91 of April 1, 1999 confirmed the principle of citizens' free choice and awareness with regards the donation of their own organs and tissue. Many studies have shown that information/communication programs aimed at secondary school students are able to significantly increase their propensity toward donation. Students who are made more aware of the topics of donation and transplantation are, in turn, a means of information for their families and acquaintances. METHODS: To be effective, communication must contain both cognitive and emotional elements. This is the principle underlying the development of an awareness-building project that involves the viewing of a play-"Two of Hearts"-telling the story of a transplant patient, followed by a brief scientific presentation aimed at clarifying the basic concepts of transplantation medicine, and then an open discussion between the students and experts. This initiative was promoted by the Piedmont Regional Tissue and Organ Procurement Coordination Agency, the Communication Office of the Piedmont Region, and the Regional Education Office for Piedmont. RESULTS: Seven meetings took place in the main town of each of the provinces that make up the Piedmont Region. These were attended by a total of 1373 students, who were each given an anonymous questionnaire. In all, 785 returned the questionnaire; of these, 95% affirmed that the contents of the meeting had been clearly presented, and 57% stated that they had spoken about it with their family and friends. The project proved be a good tool for communicating with young people and, via them, with their families; at least 2300 people were reached in this way.


Assuntos
Conscientização , Estudantes/psicologia , Obtenção de Tecidos e Órgãos , Adolescente , Humanos , Estados Unidos
4.
Transplant Proc ; 45(7): 2587-90, 2013 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-24033997

RESUMO

The culture of organ and tissue donation is not particularly well established among the general population, who often receive incorrect, incomplete information. Unfortunately, even among health workers not directly involved in the field of transplantation (laypersons), there is a poor level of knowledge concerning the underlying principles, mechanisms, and results. To increase lay health workers' knowledge and awareness of the importance of donation and transplantation, we organized an educational session for (nurses, doctors, healthcare workers, technicians, and other professionals) at a hospital coordinating service in Turin. The project was divided into 3 phases: first (February 2010), we performed an initial survey using an instrument containing 18 questions. We sought to assess the level of awareness of hospital personnel. Among 880 distributed questionnaires, 346 were compiled and returned to the authors (39.31%). During the second phase, covering the following 24 months, we held 15 educational courses on the subject of transplantation for 483 participants. In the third and last phase (February 2012), we performed a second survey, distributing, 785 questionnaires identical to the previous one, among which 404 were compiled and returned (51.46%).


Assuntos
Hospitais , Recursos Humanos em Hospital/educação , Obtenção de Tecidos e Órgãos , Humanos , Itália
5.
Transplant Proc ; 45(7): 2591-3, 2013 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-24033998

RESUMO

BACKGROUND: Over the last 20 years, an increasing number of people have immigrated into Italy. On January 1, 2011, there were 4,570,317 foreign residents in Italy, including 398,910 in the Piedmont region (Italian National Statistics Institute data). The Romanian community was the largest, followed by the Moroccan and Albanian ones. The numbers are even layer if we consider illegal immigrants. As a result, the number of foreign people with brain death has increased, as well as the number of foreign people needing organ transplantations. METHODS: The Piedmont Regional Coordination Agency of Tissue and Organ Procurement has analyzed our 1781 brain death situations between January 2004 and December 2011, including 126 (7%) in people not born in Italy (migrants). RESULTS: Non-refusal for donation occurred in 79 cases (62.7%). The agency also analyzed each community looking at number of brain deaths and non-refusals of donation. We utilized 61 donors, for 174 transplantations. Migrants in Italy are not only potential tissue and organ donors, but also potential tissue and organ recipients. Between 2004 and 2011, 222 patients who had not been born in Italy had been transplanted with: a liver (n = 66), a kidney (n = 130), a heart (n = 21) or a lung (n = 5).


Assuntos
Transplante de Órgãos , Obtenção de Tecidos e Órgãos , Migrantes , Humanos , Itália
6.
Euro Surveill ; 16(10)2011 Mar 10.
Artigo em Inglês | MEDLINE | ID: mdl-21435323

RESUMO

Following reports of West Nile neuroinvasive disease in the north-eastern area of Italy in 2009, all blood donations dating from the period between 1 August and 31 October 2009 in the Rovigo province of the Veneto region were routinely checked to exclude those with a positive nucleic acid test for West Nile virus (WNV). Only one of 5,726 blood donations was positive (17.5 per 100,000 donations; 95% confidence interval (CI): 0.4­97.3). In addition, a selection of 2,507 blood donations collected during the period from 20 July to 15 November 2009 were screened by ELISA for IgG and IgM antibodies against WNV. A positive result was received for 94 of them. The positive sera were further evaluated using immunofluorescence and plaque reduction neutralisation test (PRNT), in which only 17 sera were confirmed positive. This corresponds to a prevalence of 6.8 per 1,000 sera (95% CI: 4.0­10.9). In a case-control study that matched each of the 17 PRNT-positive sera with four negative sera with the same date of donation and same donation centre, we did not find a significant association with age and sex of the donor; donors who worked mainly outdoors were significantly more at risk to have a positive PRNT for WNV.


Assuntos
Anticorpos Antivirais/sangue , Doadores de Sangue , Imunoglobulina G/sangue , Imunoglobulina M/sangue , Febre do Nilo Ocidental/epidemiologia , Vírus do Nilo Ocidental/isolamento & purificação , Adulto , Idoso , Anticorpos Antivirais/imunologia , Ensaio de Imunoadsorção Enzimática , Feminino , Imunofluorescência , Técnica de Placa Hemolítica , Humanos , Imunoglobulina G/imunologia , Imunoglobulina M/imunologia , Incidência , Itália/epidemiologia , Masculino , Pessoa de Meia-Idade , Testes de Neutralização , Prevalência , Estudos Retrospectivos , Sensibilidade e Especificidade , Estudos Soroepidemiológicos , Febre do Nilo Ocidental/diagnóstico , Febre do Nilo Ocidental/virologia , Vírus do Nilo Ocidental/imunologia
7.
J Neurochem ; 104(1): 279-86, 2008 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-18005343

RESUMO

Brain-derived neurotrophic factor (BDNF), a member of neurotrophin family, enhances synaptic transmission and regulates neuronal proliferation and survival. Both BDNF and its tyrosine kinase receptors (TrkB) are highly expressed in the hippocampus, where an interaction with adenosine A(2A) receptors (A(2A)Rs) has been recently reported. In the present paper, we evaluated the role of A(2A)Rs in mediating functional effects of BDNF in hippocampus using A(2A)R knock-out (KO) mice. In hippocampal slices from WT mice, application of BDNF (10 ng/mL) increased the slope of excitatory post-synaptic field potentials (fEPSPs), an index of synaptic facilitation. This increase of fEPSP slope was abolished by the selective A(2A) antagonist ZM 241385. Similarly, genetic deletion of the A(2A)Rs abolished BDNF-induced increase of the fEPSP slope in slices from A(2A)R KO mice The reduced functional ability of BDNF in A(2A)R KO mice was correlated with the reduction in hippocampal BDNF levels. In agreement, the pharmacological blockade of A(2)Rs by systemic ZM 241385 significantly reduced BDNF levels in the hippocampus of normal mice. These results indicate that the tonic activation of A(2A)Rs is required for BDNF-induced potentiation of synaptic transmission and for sustaining a normal BDNF tone in the hippocampus.


Assuntos
Fator Neurotrófico Derivado do Encéfalo/metabolismo , Fator Neurotrófico Derivado do Encéfalo/farmacologia , Hipocampo/fisiologia , Receptor A2A de Adenosina/fisiologia , Transmissão Sináptica/efeitos dos fármacos , Antagonistas do Receptor A2 de Adenosina , Animais , Relação Dose-Resposta a Droga , Interações Medicamentosas , Ensaio de Imunoadsorção Enzimática/métodos , Potenciais Pós-Sinápticos Excitadores/efeitos dos fármacos , Potenciais Pós-Sinápticos Excitadores/fisiologia , Regulação da Expressão Gênica/efeitos dos fármacos , Regulação da Expressão Gênica/genética , Técnicas In Vitro , Camundongos , Camundongos Knockout , Técnicas de Patch-Clamp/métodos , Receptor A2A de Adenosina/deficiência , Triazinas/farmacologia , Triazóis/farmacologia
8.
Neurobiol Dis ; 28(2): 197-205, 2007 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-17720507

RESUMO

The effect of chronic treatment with the selective adenosine A2A receptor antagonist SCH 58261 on the behavioral and electrophysiological alterations typical of R6/2 mice (a transgenic mouse model of Huntington's disease, HD), has been studied. Starting from 5 weeks of age, R6/2 and wild type (WT) mice were treated daily with SCH 58261 (0.01 mg/kg i.p.) for 7 days. In the following weeks, the ability of mice to perform in the rotarod, plus maze and open field tests were evaluated. In addition, with electrophysiological experiments in corticostriatal slices we tested whether the well-known increased NMDA vulnerability of R6/2 mice was prevented by SCH 58261 treatment. We found that chronic treatment with SCH 58262: i) fully prevented the alterations in emotional/anxious responses displayed by R6/2 mice; ii) did not prevent the impairment in motor coordination; iii) abolished the increase in NMDA-induced toxicity observed in the striatum of HD mice. On balance, targeting A2A receptors seems to have some beneficial effects in HD even though, given the complexity of A2A receptor pharmacology and HD pathogenesis, further studies are necessary to clarify whether A2A receptor antagonists have therapeutic potential in HD.


Assuntos
Antagonistas do Receptor A2 de Adenosina , Encéfalo/efeitos dos fármacos , Doença de Huntington/tratamento farmacológico , Fármacos Neuroprotetores/farmacologia , Pirimidinas/farmacologia , Triazóis/farmacologia , Animais , Comportamento Animal/efeitos dos fármacos , Comportamento Animal/fisiologia , Encéfalo/metabolismo , Encéfalo/fisiopatologia , Corpo Estriado/efeitos dos fármacos , Corpo Estriado/metabolismo , Corpo Estriado/fisiopatologia , Modelos Animais de Doenças , Agonistas de Aminoácidos Excitatórios/toxicidade , Ácido Glutâmico/metabolismo , Doença de Huntington/metabolismo , Doença de Huntington/fisiopatologia , Deficiências da Aprendizagem/tratamento farmacológico , Deficiências da Aprendizagem/etiologia , Aprendizagem em Labirinto/efeitos dos fármacos , Aprendizagem em Labirinto/fisiologia , Transtornos Mentais/tratamento farmacológico , Transtornos Mentais/etiologia , Camundongos , Camundongos Transgênicos , Atividade Motora/efeitos dos fármacos , Atividade Motora/fisiologia , Técnicas de Cultura de Órgãos , Receptor A2A de Adenosina/metabolismo
9.
Purinergic Signal ; 3(4): 333-8, 2007 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-18404446

RESUMO

Brain-derived neurotrophic factor (BDNF), a member of the neurotrophin family, enhances synaptic transmission and regulates neuronal proliferation and survival. Functional interactions between adenosine A(2A) receptors (A(2A)Rs) and BDNF have been recently reported. In this article, we report some recent findings from our group showing that A(2A)Rs regulate both BDNF functions and levels in the brain. Whereas BDNF (10 ng/ml) increased the slope of excitatory postsynaptic field potentials (fEPSPs) in hippocampal slices from wild-type (WT) mice, it was completely ineffective in slices taken from A(2A)R knock-out (KO) mice. Furthermore, enzyme immunoassay studies showed a significant reduction in hippocampal BDNF levels in A(2A)R KO vs. WT mice. Having found an even marked reduction in the striatum of A(2A)R KO mice, and as both BDNF and A(2A)Rs have been implicated in the pathogenesis of Huntington's disease (HD), an inherited striatal neurodegenerative disease, we then evaluated whether the pharmacological blockade of A(2A)Rs could influence striatal levels of BDNF in an experimental model of HD-like striatal degeneration (quinolinic acid-lesioned rats) and in a transgenic mice model of HD (R6/2 mice). In both QA-lesioned rats and early symptomatic R6/2 mice (8 weeks), the systemic administration of the A(2A)R antagonist SCH58261 significantly reduced striatal BDNF levels. These results indicate that the presence and the tonic activation of A(2A)Rs are necessary to allow BDNF-induced potentiation of synaptic transmission and to sustain a normal BDNF tone. The possible functional consequences of reducing striatal BDNF levels in HD models need further investigation.

10.
J Neurochem ; 90(5): 1276-9, 2004 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-15312183

RESUMO

The metabotropic glutamate receptors 5 (mGlu5Rs) and the adenosine A2A receptors (A2ARs) have been reported to functionally interact in the striatum. The aim of the present work was to verify the hypothesis that the state of activation of A2A Rs could influence mGlu5R-mediated effects in the striatum. In electrophysiological experiments (extracellular recording in rat corticostriatal slices), the ability of the selective mGlu5R agonist CHPG to potentiate the reduction of the field potential amplitude induced by NMDA was prevented not only by the selective mGlu5R antagonist MPEP, but also by the selective A2AR antagonist ZM 241385. Analogously, the application of CHPG potentiated NMDA-induced toxicity (measured by LDH release) in cultured striatal neurons, an effect that was abolished by both MPEP and ZM 241385. Finally, the A2AR agonist CGS 21680 potentiated CHGP effects, an action that was reproduced and abolished, respectively, by forskolin (an activator of the cAMP/protein kinase A, PKA, pathway) and KT 5720 (a PKA inhibitor). The results indicate that A2ARs exert a permissive role on mGlu5R-induced effects in the striatum. Such an interaction may represent an additional target for the development of therapeutic strategies towards striatal disorders.


Assuntos
Adenosina/análogos & derivados , Corpo Estriado/fisiologia , Glicina/análogos & derivados , Receptores A2 de Adenosina/fisiologia , Receptores de Glutamato Metabotrópico/fisiologia , Adenosina/farmacologia , Animais , Carbazóis/farmacologia , Células Cultivadas , Corpo Estriado/efeitos dos fármacos , Relação Dose-Resposta a Droga , Interações Medicamentosas , Eletrofisiologia/métodos , Embrião de Mamíferos , Inibidores Enzimáticos/farmacologia , Agonistas de Aminoácidos Excitatórios/farmacologia , Antagonistas de Aminoácidos Excitatórios/farmacologia , Feminino , Glicina/farmacologia , Técnicas In Vitro , Indóis/farmacologia , Masculino , Potenciais da Membrana/efeitos dos fármacos , Potenciais da Membrana/fisiologia , N-Metilaspartato/farmacologia , Fenetilaminas/farmacologia , Fenilacetatos/farmacologia , Gravidez , Agonistas do Receptor Purinérgico P1 , Piridinas/farmacologia , Pirróis/farmacologia , Ratos , Ratos Wistar , Receptor de Glutamato Metabotrópico 5 , Triazinas/farmacologia , Triazóis/farmacologia
11.
Neurology ; 61(11 Suppl 6): S69-71, 2003 Dec 09.
Artigo em Inglês | MEDLINE | ID: mdl-14663014

RESUMO

Because an increased glutamate outflow is thought to play a crucial role in triggering excitotoxic neuronal death, drugs able to regulate glutamate release could be effective for the management of neurodegenerative diseases. In this article, the authors discuss the hypothesis that adenosine A2A receptor antagonists (A2A antagonists) may belong to the aforementioned category. In rats bilaterally lesioned with the excitotoxin quinolinic acid (QA) in the striatum, the A2A antagonist SCH 58261 significantly reduced the motor, EEG, and neuropathologic changes induced by the lesion. Such effects of SCH 58261 occurred only at low doses and were paralleled by an inhibition of QA-stimulated glutamate release. The role played by A2A antagonists in the regulation of glutamate outflow was also confirmed by preliminary results obtained in the model of paired-pulse stimulation in corticostriatal slices. Conversely, based on data obtained in cultured striatal neurons, A2A antagonists appear unable to directly inhibit NMDA effects. In conclusion, A2A antagonists show clear neuroprotective effects in models of brain injury, although their actual therapeutic potential needs to be confirmed in a wider range of doses and in models of neurodegenerative diseases in which presynaptic and postsynaptic effects play different relative roles.


Assuntos
Ácido Glutâmico/metabolismo , Neurotoxinas/metabolismo , Receptor A2A de Adenosina/metabolismo , Antagonistas do Receptor A2 de Adenosina , Animais , Células Cultivadas , Corpo Estriado/citologia , Corpo Estriado/efeitos dos fármacos , Corpo Estriado/metabolismo , Vias de Administração de Medicamentos , Estimulação Elétrica , Eletroencefalografia/efeitos dos fármacos , Antagonistas de Aminoácidos Excitatórios/farmacologia , Ácido Glutâmico/toxicidade , Técnicas In Vitro , Microdiálise , Atividade Motora/efeitos dos fármacos , Neurônios/efeitos dos fármacos , Neurônios/metabolismo , Fármacos Neuroprotetores/farmacologia , Neurotoxinas/toxicidade , Pirimidinas/farmacologia , Ácido Quinolínico/metabolismo , Ácido Quinolínico/toxicidade , Ratos , Receptores de N-Metil-D-Aspartato/antagonistas & inibidores , Sinapses/efeitos dos fármacos , Sinapses/metabolismo , Triazóis/farmacologia
12.
Phys Rev Lett ; 88(6): 062301, 2002 Feb 11.
Artigo em Inglês | MEDLINE | ID: mdl-11863798

RESUMO

E896 has measured Lambda production in 11.6A GeV/c Au-Au collisions over virtually the whole rapidity phase space. The midrapidity p(t) distributions have been measured for the first time at this energy and appear to indicate that the Lambda hyperons have different freeze-out conditions than protons. A comparison with the relativistic quantum molecular dynamics model shows that while there is good shape agreement at high rapidity the model predicts significantly different slopes of the m(t) spectra at midrapidity. The data, where overlap occurs, are consistent with previously reported measurements.

13.
J Neurochem ; 78(2): 349-57, 2001 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-11461970

RESUMO

In the presence of somatostatin-14 or some of its receptorial agonists, the uptake of large neutral amino acids by isolated brain microvessels was found to be inhibited up to 50%, no other transport system being affected. Although the luminal and abluminal sides of brain endothelial cells are both capable of taking up large neutral amino acids, only uptake from the abluminal side appears to be inhibited by somatostatin. The involvement of a type-2 somatostatin receptor was suggested by assays with a series of receptor-specific somatostatin agonists, and was confirmed by the release of inhibition caused by a specific type-2 receptor antagonist. A type-2-specific mRNA was indeed shown to be present in both bovine brain microvessels ex vivo and primary cultures of endothelial cells from rat brain microvessels.


Assuntos
Aminoácidos Neutros/metabolismo , Córtex Cerebral/irrigação sanguínea , Circulação Cerebrovascular/fisiologia , Endotélio Vascular/metabolismo , Microcirculação/metabolismo , Receptores de Somatostatina/fisiologia , Somatostatina/farmacologia , Animais , Transporte Biológico/efeitos dos fármacos , Bovinos , Células Cultivadas , Circulação Cerebrovascular/efeitos dos fármacos , Cinética , Microcirculação/efeitos dos fármacos , Octreotida/farmacologia , RNA Mensageiro/análise , RNA Mensageiro/genética , Receptores de Somatostatina/efeitos dos fármacos , Receptores de Somatostatina/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Transcrição Gênica
14.
Neuroreport ; 11(13): 3033-8, 2000 Sep 11.
Artigo em Inglês | MEDLINE | ID: mdl-11006989

RESUMO

In order to verify whether striatal group I metabotropic glutamate (mGlu) receptors undergo functional alteration in ageing, the effects induced by the selective agonist 3,5-dihydroxyphenylglycine (DHPG) in the striatum of young (3 months) and aged (24-25 months old) rats were compared. The ability of DHPG to stimulate phosphoinositide (PI) hydrolysis (striatal slices), to influence striatal dopamine release (in vivo microdialysis) and to potentiate the effects of NMDA on extracellular field potential amplitude (extracellular recordings on striatal slices) was reduced in the striatum of old vs young rats. These results show an age-dependent reduction in the functional response of striatal group I mGlu receptors, which may be one of the factors underlying the reduced ability aged striatum to integrate information.


Assuntos
Envelhecimento/metabolismo , Neostriado/metabolismo , Neurônios/metabolismo , Receptores de Glutamato Metabotrópico/metabolismo , Animais , Dopamina/metabolismo , Relação Dose-Resposta a Droga , Antagonistas de Aminoácidos Excitatórios/farmacologia , Glicina/análogos & derivados , Glicina/farmacologia , Hidrólise/efeitos dos fármacos , Masculino , Microdiálise/estatística & dados numéricos , N-Metilaspartato/farmacologia , Neostriado/efeitos dos fármacos , Neostriado/ultraestrutura , Neurônios/efeitos dos fármacos , Neurônios/ultraestrutura , Fosfatidilinositóis/metabolismo , Ratos , Ratos Sprague-Dawley , Receptores de Glutamato Metabotrópico/agonistas , Resorcinóis/farmacologia
15.
Am J Clin Pathol ; 110(2): 178-83, 1998 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-9704616

RESUMO

EDTA-dependent pseudothrombocytopenia (PTCP) consists of an inappropriate low platelet count caused by autoantibodies present in the serum samples reacting with platelets only in EDTA-anticoagulated blood. By using immunoprecipitation and Western blot techniques, we studied the immunochemical specificity of platelet agglutinating autoantibodies in the serum samples of 10 patients with PTCP. Furthermore, to evaluate a possible role of PTCP-associated IgG autoantibodies in increased platelet turnover, we assayed the plasma glycocalicin (GC) level and calculated the GC index for every patient. Our results provide direct evidence that an epitope located on platelet membrane glycoprotein IIb is recognized by PTCP-associated IgG antibodies; moreover GC levels in patients with EDTA-dependent PTCP were similar to control levels, thus excluding an increased platelet turnover. We conclude that antiplatelet antibodies directed against platelet cryptantigens are unlikely to have a major role in the increased removal of cells from circulation.


Assuntos
Autoanticorpos/imunologia , Plaquetas/imunologia , Ácido Edético , Imunoglobulina G/imunologia , Complexo Glicoproteico GPIIb-IIIa de Plaquetas/metabolismo , Trombocitopenia/sangue , Trombocitopenia/induzido quimicamente , Anticorpos Monoclonais/imunologia , Western Blotting , Imunofluorescência , Humanos , Contagem de Plaquetas , Complexo Glicoproteico GPIb-IX de Plaquetas/análise , Testes de Precipitina
16.
Clin Lab Haematol ; 19(3): 209-11, 1997 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-9352147

RESUMO

When studying autoantibody specificity by the indirect antiglobulin test with column agglutination techniques ether and xylene elution techniques result in haemoglobin stained eluates which give a red colouration to the gel or glass beads and do not allow the identification of positive reactions. Xylene eluates were incubated with commercially available group O-test red cell panels at 37 degrees C for 45 min in the wells of a microtitre plate in a 3:1 eluate:red cell ratio. After washing with normal saline, sensitized red cells, resuspended in low ionic strength solution (LISS), were applied onto the microtubes containing the antiglobulin serum and positive reactions were recorded after centrifugation. We studied the specificity of 35 autoantibody containing eluates from 12 patients with lymphoproliferative disorders (six having autoimmune haemolysis) and 23 HIV patients without autoimmune haemolysis. All patients had a gel or column positive (IgG) direct antiglobulin test while the tube direct antiglobulin test failed to show red cell bound IgG. We found a reactive indirect antiglobulin test in 20/23 eluates from HIV infected patients (with a panreactive specificity), in all patients with autoimmune haemolysis (one with anti-C, two with anti-E, one with anti-K and two with a panreactive specificity) and in all patients with positive direct antiglobulin test but without immune mediate haemolysis (in all cases with panreactive specificity). The method proposed is a promising tool for the study of the specificity of antibody containing haemoglobin stained eluates; in this study it allowed us to confirm that some HIV patients have specific binding of IgG on their RBC and to identify the specificity of tube test non-reactive eluates.


Assuntos
Testes de Aglutinação , Autoanticorpos/sangue , Hemoglobinas , Especificidade de Anticorpos , Teste de Coombs , Infecções por HIV/imunologia , Humanos , Transtornos Linfoproliferativos/imunologia , Sensibilidade e Especificidade
17.
Br J Pharmacol ; 120(6): 989-94, 1997 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-9134207

RESUMO

1. The density and affinity of binding sites for the delta-selective opioid ligands [3H]-[D-Ala2, Asp4]deltorphin (DELT-I), [3H]-[D-Ala2Glu4]-deltorphin (DELT-II), [3H]-[D-Pen2,D-Pen5]enkephalin (DPDPE), and [3H]-naltrindole (NTI) were determined in whole brain from 10, 15, 25 and 60 day-old C57BL mice. 2. At all ages, the analyses of the homologous displacement curves, gave best fits to single rather than to multiple site models. The binding capacity (Bmax) labelled by [3H]-NTI was about one half that labelled by [3H]-DELT-I, [3H]-DELT-II and [3H]-DPDPE. In 25 and 60 day-old mouse brain the DPDPE Bmax was 25% less than the deltorphin-II Bmax. 3. In saturation experiments, specific binding of [3H]-DELT-I on adult mouse brain homogenates was best fitted by a two-site model (34%, high affinity site, Kd = 1.08 nM and 66% low affinity sites, Kd = 39.9 nM). 4. DPDPE produced a biphasic inhibition of specific [3H]-DELTI-I binding, from 15 days of age onwards. The relative percentage of high and low affinity sites was 72% and 28% in 15 day-, 65% and 35% in 25 day- and 30% and 70% in 60 day-old mice. 5. In adult mouse brain labelled with [3H]-DELT-I, DELT-II recognized 71% of high-affinity and 29% of low-affinity sites DELT-I and DPDPE produced monophasic inhibition of specific [3H]-DELT-II binding to brain homogenates of adult mice. 6. These data suggest that a sub-population of delta-sites (probably the delta 2-subtype), recognized by DELT-I, with high affinity for DELT-II and low affinity for DPDPE develops from 25 days onward. 7. In electrically stimulated mouse vas deferens (MVD) the rank order of potency of the three delta-agonists was: DELT-I > DELT-II > DPDPE in 10 day-old mice: and DELT-I- DELT-II > DPDPE, from 25 days onward. During this time, the potency of DELT-II increased about 15 fold whereas the potency of DELT-I and DPDPE increased only 5 times. The higher efficacy of DELT-II could depend on receptor maturation towards the delta 2-subtype.


Assuntos
Encéfalo/crescimento & desenvolvimento , Receptores Opioides delta/metabolismo , Ducto Deferente/crescimento & desenvolvimento , Animais , Animais Lactentes , Encéfalo/metabolismo , Estimulação Elétrica , D-Penicilina (2,5)-Encefalina , Encefalinas/metabolismo , Feminino , Técnicas In Vitro , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Oligopeptídeos/metabolismo , Ducto Deferente/efeitos dos fármacos
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