RESUMO
DiGeorge syndrome is a disorder caused by a microdeletion on the long arm of chromosome 22. Approximately 1% of patients diagnosed with DiGeorge syndrome may have an absence of a functional thymus, which characterizes the complete form of the syndrome. These patients require urgent treatment to reconstitute T cell immunity. Thymus transplantation is a promising investigational procedure for reconstitution of thymic function in infants with congenital athymia. Here, we demonstrate a possible optimization of the preparation of thymus slices for transplantation through prior depletion of thymocytes and leukocyte cell lineages followed by cryopreservation with cryoprotective media (5% dextran FP 40, 5% Me2SO, and 5% FBS) while preserving tissue architecture. Thymus fragments were stored in liquid nitrogen at -196°C for 30 days or one year. The tissue architecture of the fragments was preserved, including the distinction between medullary thymic epithelial cells (TECs), cortical TECs, and Hassall bodies. Moreover, depleted thymus fragments cryopreserved for one year were recolonized by intrathymic injections of 3×106 thymocytes per mL, demonstrating the capability of these fragments to support T cell development. Thus, this technique opens up the possibility of freezing and storing large volumes of thymus tissue for immediate transplantation into patients with DiGeorge syndrome or atypical (Omenn-like) phenotype.
Assuntos
Síndrome de DiGeorge , Síndromes de Imunodeficiência , Humanos , Timócitos , Síndrome de DiGeorge/terapia , Timo , Células EpiteliaisRESUMO
DiGeorge syndrome is a disorder caused by a microdeletion on the long arm of chromosome 22. Approximately 1% of patients diagnosed with DiGeorge syndrome may have an absence of a functional thymus, which characterizes the complete form of the syndrome. These patients require urgent treatment to reconstitute T cell immunity. Thymus transplantation is a promising investigational procedure for reconstitution of thymic function in infants with congenital athymia. Here, we demonstrate a possible optimization of the preparation of thymus slices for transplantation through prior depletion of thymocytes and leukocyte cell lineages followed by cryopreservation with cryoprotective media (5% dextran FP 40, 5% Me2SO, and 5% FBS) while preserving tissue architecture. Thymus fragments were stored in liquid nitrogen at -196°C for 30 days or one year. The tissue architecture of the fragments was preserved, including the distinction between medullary thymic epithelial cells (TECs), cortical TECs, and Hassall bodies. Moreover, depleted thymus fragments cryopreserved for one year were recolonized by intrathymic injections of 3×106 thymocytes per mL, demonstrating the capability of these fragments to support T cell development. Thus, this technique opens up the possibility of freezing and storing large volumes of thymus tissue for immediate transplantation into patients with DiGeorge syndrome or atypical (Omenn-like) phenotype.
RESUMO
Melanoma tumor growth and progression are highly dependent on adequate blood supply through angiogenesis. Since several genes involved in angiogenesis revealed potential binding sites for the transcription factor Sp1, we have examined the effects of local inoculation of Sp1 decoy oligodeoxynucleotides (ODNs) on the growth of transplanted murine melanoma tumors and the expression of VEGF and TNF-alpha within these tumors. Treatment with Sp1 decoy ODNs, but not their mutated form, led to a significant increase (P=0.041) of the tumor necrotic area, as evaluated morphometrically. Tumor necrosis was associated with a significant decrease of microvascular density (P=0.012) and relative vascular area (P=0.026), as determined by counting CD34-positive vascular structures within the tumor microenvironment of Sp1 decoy ODNs and control ODN-treated tumors. RT-PCR experiments showed a strong decrease in the levels of VEGF188 and VEGF164 isoforms and a moderate decrease of TNF-alpha in Sp1 decoy-treated tumors. Taken together, our results indicate that Sp1 decoy ODNs may inhibit angiogenesis by affecting the gene expression of key players in angiogenesis such as TNF-alpha and VEGF. These findings indicate that Sp1 decoy ODNs may be a potential new therapeutic tool in antiangiogenic therapy.
Assuntos
Terapia Genética/métodos , Melanoma/terapia , Neovascularização Patológica/terapia , Oligodesoxirribonucleotídeos Antissenso/administração & dosagem , Neoplasias Cutâneas/terapia , Fator de Transcrição Sp1/genética , Animais , Regulação da Expressão Gênica , Melanoma/metabolismo , Melanoma/patologia , Camundongos , Camundongos Endogâmicos C57BL , Necrose , Transplante de Neoplasias , Neovascularização Patológica/metabolismo , Neovascularização Patológica/patologia , Neoplasias Cutâneas/metabolismo , Neoplasias Cutâneas/patologia , Fator de Necrose Tumoral alfa/genética , Fator A de Crescimento do Endotélio Vascular/genéticaRESUMO
We retrospectively analyzed the epidemiological features of 164 out-clinic patients with a first-onset stroke between 15 and 49 years old. Ischemic stroke occurred in 141 patients, hemorrhagic stroke in 16 patients, and venous thrombosis in 7 patients. Forty-eight percent of ischemic strokes were atherothrombotic, but no etiology was found in 32% of patients with ischemic stroke. Systemic arterial hypertension was the most frequent etiology in the hemorrhagic stroke group. The most frequent risk factors were systemic arterial hypertension, smoking, hypercholesterolemia, alcoholism and diabetes mellitus. Although stroke in young adults deserves some specific etiological investigation, we found that ordinary risk factors such as hypertension, tobacco use, hypercholesteremia and diabetes were prevalent in our population. It seems that prevention campaigns should be the target of our work.
Assuntos
Acidente Vascular Cerebral/epidemiologia , Adolescente , Adulto , Brasil/epidemiologia , Feminino , Humanos , Embolia e Trombose Intracraniana/complicações , Masculino , Pessoa de Meia-Idade , Prevalência , Estudos Retrospectivos , Fatores de Risco , Acidente Vascular Cerebral/etiologiaRESUMO
The use of computed axial tomography (CT) in the investigation of neurologic disorders is attractive for its disponibility in the health services. However, the indications of the exam and the correlation with the clinical features has not been frequently studied. We study correlation between the requests of CT and the findings reported by the radiologist, in 367 exams performed from 07/1995 to 07/ 1996. The mean age was 31.7 +/- 22.9 years. The CT were requested in decrescent order of frequency by the Services of Neurology (36.2%), Emergency room (17.4%), Pediatric Neurology (16.9%) and Internal Medicine (5.9%). The CT was more indicated in cases of seizures (30%), headache (26.2%), motor impairment (20.2%) and reduction of conscience level (16.9%). The main hypothetic diagnosis were "to discard anatomic lesions" (9.0%), not specified stroke (8.2%) and neurocisticercosis (8.2%). The result of the CT was normal in 50.4% of the exams specially those requested in cases of headache (94.4%), seizures (71.4%) and "to discard anatomic lesions"(66.7%). The more frequently CT abnormalities were hydrocephalus (5.4%), ischemic stroke (5.4%) and neoplasm (3.5%) The greatest rates of correlation were among those to discard anatomic lesions (66,7%), hydrocephalus (50%), ischemic stroke (50%) and hematoma (50%). We concluded that CT is more helpful if more clinical data is provided in the request form, so aiding the radiologist in the final report.
Assuntos
Doenças do Sistema Nervoso/diagnóstico por imagem , Tomografia Computadorizada por Raios X/estatística & dados numéricos , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Criança , Pré-Escolar , Estudos Transversais , Feminino , Cefaleia/diagnóstico por imagem , Humanos , Lactente , Masculino , Pessoa de Meia-Idade , Transtornos dos Movimentos/diagnóstico por imagem , Serviço Hospitalar de Radiologia , Convulsões/diagnóstico por imagemRESUMO
We report three patients who collectively have very representative clinical forms of neuro-Behçet and different neurological findings. The first case, male, 49 years old, presents symptoms similar to multiple sclerosis. The second case, male 15 years old, presents with parenchymatous compromise and an association with antiphospholipid antibody. And the third case, female 25 years old, presents an acute meningitis. Neuro-Behçet must always be included as a differential diagnosis of neurological disorders that have any difficulties in establishing a definite diagnosis.
Assuntos
Síndrome de Behçet/diagnóstico , Encefalopatias/diagnóstico , Adulto , Anticorpos Anticitoplasma de Neutrófilos/sangue , Síndrome de Behçet/sangue , Síndrome de Behçet/líquido cefalorraquidiano , Eletroforese das Proteínas Sanguíneas , Encefalopatias/complicações , Feminino , Humanos , Imunoglobulina A/sangue , Imunoglobulina G/sangue , Imunoglobulina M/sangue , Masculino , Pessoa de Meia-Idade , Paraparesia/etiologia , Paresia/etiologia , Albumina Sérica/análiseRESUMO
Neurofibromatosis type 1 (NF1) can virtually affect any organ, presenting most frequently with "cafe au lait" spots and neurofibromas. Vasculopathy is a known complication of NF1, but cerebrovascular disease is rare. We report the case of a 51-year-old man admitted to the hospital with a history of stroke four months before admission. On physical examination, he presented various "cafe au lait" spots and cutaneous neurofibromas. Neurologic examination demonstrated right-sided facial paralysis, right-sided hemiplegia, and aphasia. Computed tomography scan of head showed hypodense areas in the basal ganglia and centrum semiovale. Radiographs of cranium and cervical spine showed basilar impression. Angiography revealed complete occlusion of both vertebral and left internal carotid arteries, and partial stenosis of the right internal carotid artery. A large network of collateral vessels was present (moyamoya syndrome). It is an uncommon case of occlusive cerebrovascular disease associated with NF1, since most cases described in the literature are in young people, and tend to spare the posterior cerebral circulation. Basilar impression associated with this case may be considered a pure coincidence, but rare cases of basilar impression and NF1 have been described.
Assuntos
Transtornos Cerebrovasculares/etiologia , Neurofibromatose 1/complicações , Platibasia/etiologia , Transtornos Cerebrovasculares/diagnóstico , Humanos , Masculino , Pessoa de Meia-Idade , Doença de Moyamoya , Neurofibromatose 1/diagnóstico , Platibasia/diagnóstico , SíndromeRESUMO
We present the case of a patient that after chronic use of anticonvulsant drugs without proven epilepsy, developed Dupuytren's and Ledderhose's diseases. We discuss the most frequent predisposing factors, and their relationship with chronic use of anticonvulsants, particularly phenobarbitone.
Assuntos
Anticonvulsivantes/efeitos adversos , Contratura de Dupuytren/induzido quimicamente , Doenças do Pé/induzido quimicamente , Adulto , Diazepam/efeitos adversos , Contratura de Dupuytren/cirurgia , Epilepsia/tratamento farmacológico , Doenças do Pé/cirurgia , Humanos , Masculino , Fenobarbital/efeitos adversos , Fenitoína/efeitos adversos , Fatores de TempoRESUMO
The purpose was to describe the main features of headache incidence in a hospital community, its frequency and the most requested medical investigation. Due to the stressful work environment, hospital is considered to hold a high-risk population. Interviews and questionnaires were utilized. Of a 1006 files, which were randomly filled out, 987 could be analyzed. Of all, 38.5% were from headache sufferers. By using a table of pain symptoms taken from the International Headache Society classification as a pattern, headaches were assigned as migraine, tension-type and other. The mean age was 31.18 and the frequency in females was higher than in males, at any type. Family occurrence in first-degree relatives was 76.8%. Frontal location, medium intensity and pulsation were the most described features. Stress was the most frequently mentioned trigger factor. A physician was consulted only by 41.3%. Cranium X-ray was the most frequently requested exam.
Assuntos
Cefaleia/epidemiologia , Pessoal de Saúde , Doenças Profissionais/epidemiologia , Adulto , Feminino , Cefaleia/classificação , Cefaleia/etiologia , Hospitais Comunitários , Humanos , Incidência , Masculino , Doenças Profissionais/classificação , Doenças Profissionais/etiologia , Estresse Fisiológico/complicações , Inquéritos e QuestionáriosRESUMO
Previous studies have examined the arrangement of regulatory elements along the apolipoprotein B (apoB) promoter region (-3067 to +940) and a promoter fragment extending from nucleotides -150 to +124 has been demonstrated to be essential for transcriptional activation of the apoB gene in hepatic and intestinal cells. It has also been shown that transcriptional activation of apoB requires a synergistic interaction between hepatic nuclear factor-4 (HNF-4) and CCAAT/enhancer-binding protein a (C/EBPa) transcription factors. Here, we have examined the hypothesis that HNF-4 factor binding to DNA may induce a DNA helix bend, thus facilitating the communication with a C/EBPa factor located one helix turn from this HNF-4 factor in the apoB promoter. A gel electrophoretic mobility shift assay using wild type double-stranded oligonucleotides or modified wild type duplex oligonucleotides with 10 nucleotides inserted between HNF-4 and C/EBPa factor motifs showed similar retarded complexes, indicating that HNF-4 and C/EBPa factors interact independently of the distance between binding sites. However, when only one base, a thymidine, was inserted at the -71 position of the apoB promoter, the complex shift was completely abolished. In conclusion, these results regarding the study of the mechanisms involving the interaction between HNF-4 and C/EBPa factors in the apoB promoter suggest that the perfect 5'-CCCTTTGGA-3' motif is needed in order to facilitate the interaction between the two factors.
Assuntos
Apolipoproteínas B , Regiões Promotoras Genéticas , Fatores de Transcrição , Sequência de Bases , Oligonucleotídeos , Fator de Transcrição AP-1RESUMO
Previous studies have examined the arrangement of regulatory elements along the apolipoprotein B (apoB) promoter region (-3067 to +940) and a promoter fragment extending from nucleotides -150 to +124 has been demonstrated to be essential for transcriptional activation of the apoB gene in hepatic and intestinal cells. It has also been shown that transcriptional activation of apoB requires a synergistic interaction between hepatic nuclear factor-4 (HNF-4) and CCAAT/enhancer-binding protein alpha (C/EBP alpha) transcription factors. Here, we have examined the hypothesis that HNF-4 factor binding to DNA may induce a DNA helix bend, thus facilitating the communication with a C/EBP alpha factor located one helix turn from this HNF-4 factor in the apoB promoter. A gel electrophoretic mobility shift assay using wild type double-stranded oligonucleotides or modified wild type duplex oligonucleotides with 10 nucleotides inserted between HNF-4 and C/EBP alpha factor motifs showed similar retarded complexes, indicating that HNF-4 and C/EBP alpha factors interact independently of the distance between binding sites. However, when only one base, a thymidine, was inserted at the -71 position of the apoB promoter, the complex shift was completely abolished. In conclusion, these results regarding the study of the mechanisms involving the interaction between HNF-4 and C/EBP alpha factors in the apoB promoter suggest that the perfect 5'-CCCTTTGGA-3' motif is needed in order to facilitate the interaction between the two factors.
Assuntos
Apolipoproteínas B , Regiões Promotoras Genéticas , Fatores de Transcrição , Sequência de Bases , Oligonucleotídeos , Fator de Transcrição AP-1RESUMO
Previous studies have shown that the regulatory element AIC of apolipoprotein A-I is recognized by both positive and negative regulators which bind to over-lapping domains. One of these activities has been designated AIC1. Competition experiments showed that AIC1 could be competed out by oligonucleotides containing the binding site of the transcription factor NFY. In the present study, DNA binding gel electrophoresis and competition assays showed that NFY and AIC1 recognized the same binding site on element AIC. This site contains a CCACT motif and differs by one residue from the consensus CCAAT binding motif of NFY. Cotransfection of HepG2 cells with both the -177 to -148 apoA-I CAT constructs and plasmid expressing NFY alpha and NFY beta, transactivated the apoA-I promoter by 1.8 fold, indicating that NFY is a positive activator of the apoA-I gene.
Assuntos
Apolipoproteína A-I/genética , Proteínas de Ligação a DNA/metabolismo , Regiões Promotoras Genéticas , Sequências Reguladoras de Ácido Nucleico , Fatores de Transcrição/metabolismo , Ativação Transcricional , Animais , Sítios de Ligação , Proteínas Estimuladoras de Ligação a CCAAT , Células COS , Humanos , Masculino , Ratos , Ratos Sprague-Dawley , Transfecção , Células Tumorais CultivadasRESUMO
Congenital generalized lipodystrophy is a rare inherited disease. One of its features is a disturbance in lipid metabolism characterized by hypercholesterolemia and hypertriglyceridemia. A brother and a sister with congenital generalized lipodystrophy, an 8-year old male and a 12-year old female were studied. The mother and a 6-year old brother were healthy. The genetic analysis of Sstl RFLP of the apo AI-CIII-AIV gene cluster showed the presence of the rare Sstl allele (S2) in the patients but not in the healthy mother and brother. As this uncommon allele has been reported to be related to high plasma triglyceride levels, this association could be relevant in explaining in part the hypertriglyceridemia observed in these patients.
Assuntos
Alelos , Apolipoproteínas/genética , Lipodistrofia/congênito , Lipodistrofia/genética , Família Multigênica/genética , Triglicerídeos/sangue , Criança , Feminino , Humanos , Masculino , Polimorfismo GenéticoRESUMO
We report the molecular characterization of a middle repetitive DNA sequence, named C6, isolated from the Trypanosoma cruzi genome. C6 appears to be a composite repeated element since 3 subregions may be defined within it on the basis of sequence similarities with other T. cruzi genomic sequences. Sequences homologous to C6 are interspersed in the genome and can be mapped out on most chromosomal bands of different T. cruzi. strains. The copy number of the C6 element is about 1000 per haploid genome. Given the species specificity and different genomic distribution of C6 homologous sequences among the T. cruzi strains the C6 element could be a useful probe for diagnosis and typing of parasites. C6 is a polymorphic marker with potential as a tool for physical mapping of the T. cruzi genome.
Assuntos
Genoma de Protozoário , Sequências Repetitivas de Ácido Nucleico , Trypanosoma cruzi/genética , Animais , Sequência de Bases , Southern Blotting , Doença de Chagas/diagnóstico , Doença de Chagas/parasitologia , DNA de Protozoário/genética , Eletroforese em Gel de Campo Pulsado , Dados de Sequência Molecular , Reação em Cadeia da Polimerase , Polimorfismo Genético/genética , Sequências Repetitivas de Ácido Nucleico/genética , Análise de Sequência de DNA , Especificidade da Espécie , Trypanosoma cruzi/isolamento & purificaçãoRESUMO
Possible associations between coronary heart disease (CHD) and restriction fragment length polymorphisms (RFLPs) in the apo AI-CII-AIV cluster and the apo B gene were investigated in a Brazilian population consisting of 46 patients with CHD and 24 individuals without evidence of CHD. A preliminary genetic analysis of SstI RFLP in the apo AI-CII-AIV cluster showed a significantly higher frequency of the rare SstI allele (S2) in CHD patients as compared with controls. No significant differences were found in the frequencies of PstI RFLP in the apo AI-CII-AIV cluster or XbaI and EcoRI RFLPs in the apo B gene between CHD patients and controls. Moreover, no association was seen between the RFLPs studied and myocardial infarction or plasma cholesterol or triglyceride levels.
Assuntos
Apolipoproteína A-I/genética , Apolipoproteínas B/genética , Polimorfismo Genético/genética , Adulto , Idoso , Apolipoproteína A-I/análogos & derivados , Brasil , Doença das Coronárias/genética , Feminino , Humanos , MasculinoRESUMO
Possible associations between coronary heart disease (CHD) and restriction fragment length polymorphisms (RFLPs) in the apo AI-CIII-AIV cluster and the apo B gene were investigated in a Brazilian population consisting of 46 patients with CHD and 24 individuals without evidence of CHD. A preliminary genetic analysis of SstI RFLP in the apo AI-CIII-AIV cluster showed a significantly higher frequency of the rare SstI allele (S2) in CHD patients as compared with controls. No significant differences were found in the frequencies of PstI RFLP in the apo AI-CIII-AIV cluster or XbaI and EcoRI RFLPs in the apo B gene between CHD patients and controls. Moreover, no association was seen between the RFLPs studied and myocardial infarction or plasma cholesterol or triglyceride levels.
Assuntos
Adulto , Idoso , Feminino , Humanos , Apolipoproteína A-I/genética , Apolipoproteínas B/genética , Doença das Coronárias/genética , Polimorfismo Genético/genética , Apolipoproteína A-I/análogos & derivados , BrasilRESUMO
A fragment of Trypanosoma cruzi ribosomal intergenic spacer (IGS) located at 6.7 kb from the 3' end of the 24S rRNA gene was analyzed. This IGS fragment is characterized by the presence of three types of repetitive elements (designated Spacer Repetitive Elements, SRE), short direct repeats (5-6 bp) and chi-like recombinational sequences. SRE elements are composed of relatively short repeats (43-145 bp) which show variabilities consisting of nucleotide changes, insertions and deletions. SRE-1 element (145 bp) has a short oligo(dA) tail at the end of the repeat and can be found flanked by other SRE elements. SRE elements are species-specific, suggesting that probes based on them may be diagnostic for Trypanosoma cruzi.
Assuntos
Sequências Repetitivas de Ácido Nucleico , Trypanosoma cruzi/genética , Animais , Sequência de Bases , DNA de Protozoário/genética , DNA Ribossômico/genética , Genoma , Dados de Sequência Molecular , Mapeamento por Restrição , Especificidade da Espécie , Trypanosoma/genéticaRESUMO
It is reported a case of a 61 years-old man with progressive asthenia, disfagia, disphonia and diplopia, of variable intensity during the day, who had a very good response to anticolinesterasic drugs and corticosteroids. The repetitive stimulation tests at low frequency, resulted in large increment (maximum 275%) of the basal voltage after exercise. At high frequency he also had a large increment. Radiological and laboratory investigation three times in a seven-year period was normal, without evidence of any neoplasia. Muscle biopsy showed a type II muscle fiber atrophy. The repetitive stimulation tests repeated three times, was typical of myasthenia gravis in one occasion and in the other two, typical of myasthenic syndrome (increment of 418%). A discussion about other cases with similar findings is made after a review of the literature.
Assuntos
Eletrodiagnóstico , Doenças Musculares/diagnóstico , Miastenia Gravis/diagnóstico , Diagnóstico Diferencial , Estimulação Elétrica , Humanos , Masculino , Pessoa de Meia-Idade , Miastenia Gravis/fisiopatologia , Esforço Físico , Receptores Colinérgicos/análiseRESUMO
The genito-femoral nerves of ten albinic rats chronically intoxicated with DDT (5 mg/kg day during 180 days) was studied by teased fibers examination. It was found 85,37% types A + B fibers, 13,63% type C and 1% of types D, E, F and G. It is concluded that there occurs an axonal degeneration of myelin induced by chronic DDT intoxication.