RESUMO
BACKGROUND: The Addenbrooke's Cognitive Examination III (ACE-III), an adaptation of the ACE cognitive screening test, has been demonstrated to have high sensitivity and specificity in detecting cognitive impairment in patients with dementia and other neurological and psychiatric disorders. Although the Spanish-language version of the ACE-III has already been validated in Spain, it is yet to be validated in Latin America. The aim of this study was to validate the ACE-III test in an Argentinean and Chilean population. METHODS: ACE-III was administered to 70 patients with Alzheimer disease, 31 patients with behavioural variant frontotemporal dementia, and a control group of 139 healthy volunteers. Participants were recruited at centres in both countries. RESULTS: The Spanish-language version of ACE-III was found to have good internal consistency (Cronbach's alpha=0.87). We found significant differences in total ACE-III scores between patients with Alzheimer disease and controls (p< .05) and between patients with Alzheimer disease and bvFTD (p< .05). With a cut-off point of 86, 98.6% of AD patients, 83.9% of behavioural variant frontotemporal dementia patients, and 84.2% of controls were correctly classified. CONCLUSIONS: This study shows that the Spanish-language version of ACE-III continues to be an effective tool for detecting cognitive dysfunction in patients with dementia.
Assuntos
Doença de Alzheimer/diagnóstico , Disfunção Cognitiva/diagnóstico , Idioma , Testes de Estado Mental e Demência/normas , Tradução , Idoso , Argentina , Chile , Feminino , Demência Frontotemporal/diagnóstico , Humanos , Masculino , Reprodutibilidade dos Testes , Sensibilidade e EspecificidadeRESUMO
Several studies have reported that about 65 % of patients with relapsing-remitting multiple sclerosis (RRMS) suffer from cognitive impairment, with executive dysfunction being the most frequently described. Even if several executive screening tests have been designed to specifically detect executive deficits, few studies have investigated their ability to tackle such dysfunction particularly in multiple sclerosis (MS). The aim of the present study was to evaluate the sensitivity and specificity of the INECO frontal screening (IFS) in the detection of executive dysfunction in patients with relapsing-remitting MS (RRMS). 54 patients with RRMS were included in the study. 34 presented executive dysfunction while 20 did not. 32 control subjects matched for age, sex, and educational level were also included. All were evaluated with the IFS and with a battery of classical executive tests. A patient was considered to have executive dysfunction if he/she scored a one and a half standard deviation below the control mean in at least one of the classical executive tests. Sensitivity and specificity of the IFS in its ability to detect executive dysfunction in MS was analyzed. Using a cut-off of 25.5 points, sensitivity of the IFS was 73.53 %, and specificity 78.13 % in differentiating controls from MS patients with executive dysfunction. The IFS showed excellent concurrent validity with executive tasks. The IFS can be considered a brief, easy-to-administer, cost-less tool for the detection of executive dysfunction in patients with RRMS.
Assuntos
Função Executiva , Esclerose Múltipla Recidivante-Remitente/diagnóstico , Esclerose Múltipla Recidivante-Remitente/psicologia , Testes Neuropsicológicos , Adulto , Humanos , Curva ROC , Sensibilidade e EspecificidadeRESUMO
Live-cell imaging with fluorescent protein labeling is providing major new insights into nuclear dynamics in filamentous fungi. With this approach we provide a detailed report of nuclear organization and behavior during mitosis in the bean pathogen Colletotrichum lindemuthianum. Nuclear division and nuclear migration were analyzed in ungerminated conidia, conidial germlings and the mature colony. Ungerminated conidia were uninucleate and completion of mitosis was found not to be essential for germ tube formation, conidial anastomosis tube (CAT) formation or fusion. Nuclei in fused conidial germlings exhibited asynchronous mitoses, and nuclear migration through fused CATs occurred after the nuclei had divided. Different patterns of nuclear division were found in vegetative hyphae of the mature colony. Synchronous, parasynchronous and asynchronous patterns of mitosis were observed in apical hyphal compartments at the colony border, while only synchronous and asynchronous mitoses occurred in subapical hyphal compartments. These findings have revealed unexpected diversity in the patterns of mitosis in different cells of C. lindemuthianum.
Assuntos
Divisão do Núcleo Celular , Colletotrichum/citologia , Colletotrichum/crescimento & desenvolvimento , Mitose , Esporos Fúngicos/citologiaRESUMO
BACKGROUND: We recently demonstrated that decline in fluid intelligence is a substantial contributor to frontal deficits. For some classical 'executive' tasks, such as the Wisconsin Card Sorting Test (WCST) and Verbal Fluency, frontal deficits were entirely explained by fluid intelligence. However, on a second set of frontal tasks, deficits remained even after statistically controlling for this factor. These tasks included tests of theory of mind and multitasking. As frontal dysfunction is the most frequent cognitive deficit observed in early Parkinson's disease (PD), the present study aimed to determine the role of fluid intelligence in such deficits. METHOD: We assessed patients with PD (n=32) and control subjects (n=22) with the aforementioned frontal tests and with a test of fluid intelligence. Group performance was compared and fluid intelligence was introduced as a covariate to determine its role in frontal deficits shown by PD patients. RESULTS: In line with our previous results, scores on the WCST and Verbal Fluency were closely linked to fluid intelligence. Significant patient-control differences were eliminated or at least substantially reduced once fluid intelligence was introduced as a covariate. However, for tasks of theory of mind and multitasking, deficits remained even after fluid intelligence was statistically controlled. CONCLUSIONS: The present results suggest that clinical assessment of neuropsychological deficits in PD should include tests of fluid intelligence, together with one or more specific tasks that allow for the assessment of residual frontal deficits associated with theory of mind and multitasking.
Assuntos
Função Executiva/fisiologia , Inteligência/fisiologia , Doença de Parkinson/fisiopatologia , Idoso , Humanos , Pessoa de Meia-Idade , Teoria da Mente/fisiologiaRESUMO
It has been hypothesized that horizontal gene/chromosome transfer and parasexual recombination following hyphal fusion between different strains may contribute to the emergence of wide genetic variability in plant pathogenic and other fungi. However, the significance of vegetative (heterokaryon) incompatibility responses, which commonly result in cell death, in preventing these processes is not known. In this study, we have assessed this issue following different types of hyphal fusion during colony initiation and in the mature colony. We used vegetatively compatible and incompatible strains of the common bean pathogen Colletotrichum lindemuthianum in which nuclei were labelled with either a green or red fluorescent protein in order to microscopically monitor the fates of nuclei and heterokaryotic cells following hyphal fusion. As opposed to fusion of hyphae in mature colonies that resulted in cell death within 3 h, fusions by conidial anastomosis tubes (CAT) between two incompatible strains during colony initiation did not induce the vegetative incompatibility response. Instead, fused conidia and germlings survived and formed heterokaryotic colonies that in turn produced uninucleate conidia that germinated to form colonies with phenotypic features different to those of either parental strain. Our results demonstrate that the vegetative incompatibility response is suppressed during colony initiation in C. lindemuthianum. Thus, CAT fusion may allow asexual fungi to increase their genetic diversity, and to acquire new pathogenic traits.
Assuntos
Fungos/genética , Variação Genética , Plantas/microbiologia , Núcleo Celular , Transferência Genética Horizontal , Hifas , Esporos FúngicosRESUMO
BACKGROUND: The Addenbrooke's Cognitive Examination Revised (ACE-R) is an improved version of the earlier brief screening test which has been validated in English with high sensitivity and specificity to detect cognitive dysfunction. The aim of this study was to validate the Spanish version of the ACE-R in an Argentine population. METHODS: A group of patients with Alzheimer Disease (AD) and patients with behavioural variant Frontotemporal Dementia (bvFTD) paired by age, sex, and years of education with healthy controls were assessed using the ACE-R. Stage of dementia was measured with the Clinical Dementia Rating Scale (CDR). The English version of the ACE-R was first translated into Spanish and then back-translated into English by two blind independent experts. RESULTS: Internal reliability was very good (Cronbach's alpha=0.89). Concurrent validity, determined by the correlation between total ACE-R and CDR was significant (P<.001) and inter-rater reliability was excellent (Cohen's kappa=0.98). Controls significantly outperformed AD and bvFTD patients on most subdomains of the ACE-R, with significant differences between the dementia groups. With a cut-off score of 85 points, sensitivity was 97.5% and specificity was 88.5%, with a likelihood ratio of 99.3 for the detection of dementia. The ACE-R showed higher sensitivity than the MMSE for the detection of dementia. CONCLUSIONS: The Spanish version of the ACE-R is a brief yet reliable screening tool for the detection of early cognitive impairment and has shown to discriminate between bvFTD and AD.
Assuntos
Transtornos Cognitivos/diagnóstico , Testes Neuropsicológicos/normas , Idoso , Idoso de 80 Anos ou mais , Doença de Alzheimer/diagnóstico , Argentina , Demência Frontotemporal/diagnóstico , Humanos , Idioma , Psicometria , Curva ROC , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , EspanhaRESUMO
Fusion of conidia and conidial germlings by means of conidial anastomosis tubes (CATs) is a common phenomenon in filamentous fungi, including many plant pathogens. It has a number of different roles, and has been speculated to facilitate parasexual recombination and horizontal gene transfer between species. The bean pathogen Colletotrichum lindemuthianum naturally undergoes CAT fusion on the host surface and within asexual fruiting bodies in anthracnose lesions on its host. It has not been previously possible to analyze the whole process of CAT fusion in this or any other pathogen using live-cell imaging techniques. Here we report the development of a robust protocol for doing this with C. lindemuthianum in vitro. The percentage of conidial germination and CAT fusion was found to be dependent on culture age, media and the fungal strain used. Increased CAT fusion was correlated with reduced germ tube formation. We show time-lapse imaging of the whole process of CAT fusion in C. lindemuthianum for the first time and monitored nuclear migration through fused CATs using nuclei labelled with GFP. CAT fusion in this pathogen was found to exhibit significant differences to that in the model system Neurospora crassa. In contrast to N. crassa, CAT fusion in C. lindemuthianum is inhibited by nutrients (it only occurs in water) and the process takes considerably longer.
Assuntos
Colletotrichum/citologia , Colletotrichum/fisiologia , Fabaceae/microbiologia , Doenças das Plantas/microbiologia , Esporos Fúngicos/fisiologia , Imagem com Lapso de Tempo/métodos , Núcleo Celular/metabolismo , Colletotrichum/genética , Proteínas de Fluorescência Verde/metabolismo , Microscopia ConfocalRESUMO
INTRODUCTION: In clinical practice it is often difficult to establish whether cognitive impairment is secondary to an affective disorder or a dementing process. AIM: To describe the cognitive performance on the Spanish version of the Addenbrooke's Cognitive Examination (ACE) of patients with early dementia and depression. SUBJECTS AND METHODS: 77 patients with early dementia (53 Alzheimer disease; 24 frontotemporal dementia), 17 patients with major depression and 54 healthy volunteers were tested with the Spanish version of the ACE. RESULTS: Alzheimer disease and frontotemporal dementia groups were significantly lower than the control group and the major depression group. When the major depression group was compared with the control group no significant differences were found. CONCLUSIONS: The cognitive performance in the ACE is different in patients with early dementia and patient with depression.
Assuntos
Demência/diagnóstico , Transtorno Depressivo Maior/diagnóstico , Idoso , Diagnóstico Diferencial , Feminino , Humanos , Idioma , Masculino , Pessoa de Meia-Idade , Testes NeuropsicológicosRESUMO
Cognitive function and diffusion tensor imaging were assessed in a group of 12 patients with early relapsing-remitting multiple sclerosis (disease duration Assuntos
Transtornos Cognitivos/patologia
, Imagem de Difusão por Ressonância Magnética
, Lobo Frontal/patologia
, Esclerose Múltipla Recidivante-Remitente/patologia
, Adolescente
, Adulto
, Cognição
, Transtornos Cognitivos/etiologia
, Feminino
, Humanos
, Masculino
, Rememoração Mental
, Esclerose Múltipla Recidivante-Remitente/complicações
, Vias Neurais/patologia
, Testes Neuropsicológicos
, Aprendizagem Verbal
Assuntos
Doenças dos Gânglios da Base/etnologia , Doenças dos Gânglios da Base/genética , Predisposição Genética para Doença/genética , Proteínas do Tecido Nervoso/genética , Ataxias Espinocerebelares/etnologia , Ataxias Espinocerebelares/genética , Adulto , Idoso , Argentina/etnologia , Ataxina-10 , Gânglios da Base/fisiopatologia , Doenças dos Gânglios da Base/fisiopatologia , Cerebelo/fisiopatologia , Análise Mutacional de DNA , Feminino , Frequência do Gene , Marcadores Genéticos , Testes Genéticos , Humanos , Indígenas Sul-Americanos/etnologia , Indígenas Sul-Americanos/genética , Masculino , Pessoa de Meia-Idade , Mutação/genética , Linhagem , Ataxias Espinocerebelares/fisiopatologia , População Branca/etnologia , População Branca/genéticaRESUMO
Lethal yellowing (LY) of coconut palm (Cocos nucifera L.) caused by a subgroup 16SrIV-A phytoplasma has been present along the northern coast and adjacent Bay Islands of Honduras since 1996. In the southern municipalities of San Esteban and Guanaco, approximately 150 km from the Atlantic coast, substantial numbers of coyol palm (Acrocomia aculeata (Jacq.) Lodd. ex Mart.) and several coconut palms growing nearby were either dead or in an advanced stage of decline during 2005. Declining palms of both species exhibited loss of fruit, withered inflorescences, and dried, discolored (grayish brown) leaves. Most or all leaves on declining palms had collapsed and hung downward around the stem. Samples (3 to 5 g) of tissue were excised from stems of 20 coyol and 2 coconut palms. DNA was extracted from each sample using a previously described protocol (2) and assayed for phytoplasma DNA using polymerase chain reaction (PCR) employing phytoplasma universal rRNA primer pair P1 (1) and P7 (4). A product of expected size (approximately 1.8 kb) was amplified from 12 of 20 diseased coyol palms, 2 of 2 diseased coconut palms and from DNA of a Florida-grown Chinese fan palm (Livistona chinensis (Jacq.) R. Br. ex Mart.) with LY symptoms included as a known positive control. Amplified P1/P7 products used in a nested PCR assay with 16SrIV-specific primer pair LY16Sf (2) and LY16Sr (5'-GCTTACGCA-GTTAGGCTGTC-3') yielded a product of approximately 1.39 kb. Neither primary nor nested PCRs generated a product from DNA of healthy coconut palm. Poor quality of tissues within stems of declining palms may have contributed to lack of amplification of any product from eight coyol samples. A reevaluation of DNA samples using PCR employing nonribosomal primer pair LYF1/LYR1, which specifically detects subgroup 16SrIV-A phytoplasmas (3), failed to amplify an expected 1-kb product from any palm sample other than the LY positive control. Digestion of nested amplification products (1.39 kb) with AluI endonuclease and electrophoresis of digests through 8% polyacrylamide gels revealed the same three-fragment restriction pattern for all phytoplasmas detected in coyol and coconut palms. The pattern differed from the five-fragment profile of LY phytoplasma rDNA included for comparative purposes. These results indicate that coyol palm is a new phytoplasma host and that decline symptoms on coyol and adjacent coconuts were not a consequence of natural dispersion of subgroup 16SrIV-A phytoplasmas from the northern coast. The 16S rDNA sequences amplified from coyol palm decline (CPD) (GenBank Accession No. DQ321818) and coconut decline (CLDO) (GenBank Accession No. DQ321819) phytoplasmas were coidentical and most similar (99.87%) to that of Yucatan coconut lethal decline (LDY), a known subgroup 16SrIV-B strain. This relationship was further supported by phylogenetic analysis of rDNA sequences. On the basis of these findings, we have tentatively concluded that strains CPD and CLDO represent new members of the coconut lethal yellows subgroup 16SrIV-B. References: (1) S. Deng and C. Hiruki, J. Microbiol. Methods 14:53, 1991 (2) N. A. Harrison et al. Plant Dis. 86:676, 2002 (3) N. A. Harrison et al. Plant Pathol. 43:998, 1994. (4) C. D. Smart et al. Appl. Environ. Microbiol. 62:2988, 1996.
RESUMO
Apesar da grande preocupação mundial com as doenças parasitárias, as helmintoses continuam vencendo essa batalha. A sua disseminação aumenta com a diminuição das condições sócio-econômicas das populações. O desenvolvimento farmacotécnico deve ser feito de uma maneira racional, para que agregue em um mesmo produto qualidade e baixo custo. Este trabalho objetivou a obtenção de comprimidos de mebendazol 100mg, bem como, desenvolver estudo comparativo com medicamentos genéricos disponíveis no mercado. O desenvolvimento das formulações foi iniciado por meio de um planejamento qualitativo e quantitativo de excipientes, com lotes de bancada com até 300g. Foram produzidos nove lotes e avaliados os parâmetros: aspecto, peso médio, friabilidade, dureza, desintegração, dissolução e doseamento. Os comprimidos de mebendazol 100mg e os medicamentos genéricos avaliados apresentaram-se com resultados dentro das especificações demonstrando que existe entre eles uma equivalência frente aos parâmetros avaliados. A técnica empregada, compressão direta, exceto para o LB I (granulação via úmida), para obtenção dos comprimidos, mostrou-se adequada para produção destes, pois resultou em comprimidos dentro das especificações farmacopéicas, além de agregar vantagens industriais para a produção pela via direta.
Assuntos
Antinematódeos/farmacocinética , Mebendazol/farmacocinética , Medicamentos Genéricos/farmacocinética , Preparações FarmacêuticasRESUMO
INTRODUCTION: The Addenbrooke's Cognitive Examination (ACE) is a brief bedside test battery to detect mild dementia and differentiate frontotemporal dementia (FTD) from Alzheimer's disease (AD). AIM: To validate the ACE in Spanish. PATIENTS AND METHODS: The study evaluated the Spanish version of ACE on 128 subjects consisting in two groups a patient group (n = 76) and a control subjects group (n = 52). The patient group was divided in AD (n = 54) based on the NINCDS-ADRDA criteria and FTD (n = 22) based on the Lund y Manchester criteria. All patients underwent clinical, neuropsychological, radiologic (MRI, CT, and SPECT), and laboratory evaluations. Group's differences were evaluated using ANOVA. The internal consistency of the Spanish version of the ACE was measured using the Cronbach's alpha coefficient. The discriminative capability of the Spanish version of the ACE was examined by the receiver operating characteristic (ROC) analysis. RESULTS: The cut-off score of 86 showed a sensitivity of 92% (CI 95% = 83.6-97.0) and a specificity of 96.2% (CI 95% = 86.8-99.4). The ROC curve showed higher sensitivity and specificity of the ACE than the Mini-Mental State Examination in discriminating the dementia and control group. The VLOM ratio (verbal fluency + language)/(orientation + memory) of < 1.82 discriminated for FTD and > 4.87 discriminated for AD. CONCLUSION: The Spanish version of ACE is a brief and reliable instrument for early detection of dementia in highly educated people and offers a simple objective index to differentiate AD and FTD. More studies in less educated people are warranted.
Assuntos
Doença de Alzheimer/diagnóstico , Demência/diagnóstico , Lobo Frontal/patologia , Testes Neuropsicológicos , Idoso , Idoso de 80 Anos ou mais , Doença de Alzheimer/patologia , Doença de Alzheimer/fisiopatologia , Demência/patologia , Demência/fisiopatologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Escalas de Graduação Psiquiátrica , Curva ROC , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , EspanhaRESUMO
UNLABELLED: This study aimed to investigate the biodistribution of the 99mTc-ceftizoxime in normal rats and in rats bearing septic and sterile induced abscess. MATERIAL AND METHODS: Three groups of rats were studied. a) Six normal rats b) 15 rats with E. coli induced abscess and c) 15 rats with sterile zymosan induced abscess. Septic abscess was induced with 2 x 10(8) colony forming units of E. coli and sterile one with 0.1 mL of 5% sterile Zymosan. 24 h after the abscess induction, 12 MBq of 99mTc-CFT were injected iv. and whole body images were collected at 30 min, 1, 2, 4 and 6 h p.i. Areas of interest were drawn and lesion/background index was calculated. The 6 normal rats were scanned at the same times, killed at 6 h p.i and kidney, liver, spleen, lung, heart and muscle activity were measured. Each organ was weighed, cut and its activity measured. Parallelly, the biological activity of the labeled antibiotic and its binding to the E. coli and S. aureus bacteria were analyzed. RESULTS: High biliary excretion was seen in all rats. Organ measurement showed the maximal uptake in kidney and very low uptake in muscles. Mean +/- s.d abscess/background ratio at 30 min, 1, 2, 4 and 6 h were 2.60 +/- 0.36, 2.67 +/- 0.66, 2.6 0 +/- 0.58, 2.78 +/- 0.84, 3.24 +/- 1.00 for septic abscess and 2.37 +/- 0.39, 2.10 +/- 0.38, 1.97 +/- 0.34, 1.82 +/- 0.25, 1.65 +/- 0.23 for aseptic abscess. The 99mTc-CFT uptake was significantly higher in the septic abscess than in sterile one (p < 0.05). The 99mTc-CFT uptake in the septic abscess remains stable or increases until along the 6 h. The 99mTc-CFT uptake in the aseptic abscess decreases along the time. CONCLUSIONS: The scintigraphy with 99mTc-CFT seems able to differentiate sterile inflammation from infection. High biliary excretion limits its application in abdomen. Main application could be diagnosis of osteoarticular infection.
Assuntos
Abscesso/diagnóstico por imagem , Ceftizoxima/análogos & derivados , Compostos de Organotecnécio , Compostos Radiofarmacêuticos , Abscesso/microbiologia , Animais , Ceftizoxima/farmacocinética , Diagnóstico Diferencial , Compostos de Organotecnécio/farmacocinética , Cintilografia , Compostos Radiofarmacêuticos/farmacocinética , Ratos , Ratos Sprague-DawleyRESUMO
A patient with generalized dystonia due to Wilson's disease obtained marked improvement in response to smoking cannabis.
Assuntos
Cannabis , Distúrbios Distônicos/tratamento farmacológico , Distúrbios Distônicos/etiologia , Degeneração Hepatolenticular/complicações , Fitoterapia , Adulto , Avaliação da Deficiência , Degeneração Hepatolenticular/tratamento farmacológico , Humanos , Masculino , Preparações de Plantas/uso terapêutico , Resultado do TratamentoRESUMO
We here report on 3 patients with essential tremor, otherwise unresponsive to pharmacological treatment, who greatly benefited from low doses of topiramate (50 mg/d). No side effects were observed and improvement was sustained during a mean of 7 months (range 3-12 months) follow up. Our results suggest that topiramate titration should be performed gradually, so as not to neglect cases responsive to low doses.
Assuntos
Anticonvulsivantes/administração & dosagem , Anticonvulsivantes/uso terapêutico , Tremor Essencial/tratamento farmacológico , Frutose/análogos & derivados , Frutose/administração & dosagem , Frutose/uso terapêutico , Fármacos Neuroprotetores/administração & dosagem , Fármacos Neuroprotetores/uso terapêutico , Antagonistas Adrenérgicos beta/uso terapêutico , Idoso , Anticonvulsivantes/efeitos adversos , Transtorno Bipolar/complicações , Transtorno Bipolar/fisiopatologia , Relação Dose-Resposta a Droga , Frutose/efeitos adversos , Humanos , Masculino , Fármacos Neuroprotetores/efeitos adversos , Testes Neuropsicológicos , Primidona/uso terapêutico , Propranolol/uso terapêutico , TopiramatoRESUMO
Fibrocartilagenous embolism (FCE) of the intervertebral disc represents a very rare cause of spinal infarct. Up to now only 33 others cases in human beings have been reported in the literature, most of them diagnosed post mortem. We present a 14-year-old boy who developed acute dorsal back pain after lifting a heavy gate, followed by progressive paraparesis. An MRI of the spine showed a degenerative disc at D10-D11 without compromise of the spinal canal lumen associated with an acute Schmorl's nodule situated in the superior endplate of D11. A week later, a second MRI disclosed an intraxial spinal cord lesion at D7-D8 vertebral level involving the vascular territory of the anterior spinal artery. It also showed an abnormal signal located in the posterior third of the D8 vertebral body. These clinical and neuro-radiological findings are similar to those mentioned in the literature and support the diagnosis of an anterior spinal infarct secondary to a probable fibrocartilaginous embolism. This case highlights the importance of considering this etiology among the causes of spinal cord infarct, especially in young people, and underlines the utility of MRI in its diagnosis in vivo.
Assuntos
Embolia/complicações , Disco Intervertebral/patologia , Paraparesia/etiologia , Adolescente , Embolia/diagnóstico , Embolia/diagnóstico por imagem , Humanos , Disco Intervertebral/diagnóstico por imagem , Imageamento por Ressonância Magnética , Masculino , Paraparesia/patologia , Radiografia , Medula Espinal/diagnóstico por imagem , Medula Espinal/patologia , Coluna Vertebral/diagnóstico por imagem , Coluna Vertebral/patologia , Vértebras TorácicasRESUMO
Isolation of DNA for PCR is time-consuming and involves many reagents. The aim of this work was to optimise a rapid and easy PCR methodology without previous DNA isolation. Different strains of the phytopathogenic fungus Colletotrichum lindemuthianum were used. Protoplasts were generated using lytic enzymes under high incubation temperatures using different methodologies to obtain the template. A rapid (10 minute) methodology was successful for smaller amplicons (<750 bp).
Assuntos
Colletotrichum , Técnicas In Vitro , Reação em Cadeia da Polimerase/métodos , MétodosRESUMO
Isolation of DNA for PCR is time-consuming and involves many reagents. The aim of this work was to optimise a rapid and easy PCR methodology without previous DNA isolation. Different strains of the phytopathogenic fungus Colletotrichum lindemuthianum were used. Protoplasts were generated using lytic enzymes under high incubation temperatures using different methodologies to obtain the template. A rapid (10 minute) methodology was successful for smaller amplicons ( 750 bp).
A extração de DNA para realização de PCR é demorada e envolve vários reagentes. O objetivo deste trabalho foi desenvolver uma metodologia rápida e fácil que permita utilizar a técnica de PCR sem necessidade de extração de DNA. Diferentes isolados de Colletotrichum lindemuthianum foram utilizados. Para obtenção do DNA moldes foram testados protoplastos obtidos através do uso de enzimas líticas, incubados sob altas temperaturas e diferentes metodologias. Uma metodologia rápida (10 minutos) foi desenvolvida para produtos de PCR menores ( 750 pb).
RESUMO
The aim of this study was to retrospectively compare the diagnostic accuracy of (99m)Tc hexamethylpropyleneamine oxime (HMPAO) white blood cell scintigraphy in patients with a suspicion of active inflammatory bowel disease by using two different cell suspension media: leukocyte poor plasma (LPP) and Hanks' balanced salt solution (HBSS). Leukocytes from 30 patients were labelled using LPP and in 28 cases using HBSS. In the LPP method the leukocytes were resuspended in 0.5 ml cell-free plasma while in the HBSS method the cells were resuspended in 0.5 ml HBSS. Scintigraphic images were obtained at 30 min and 2 h after injection of 185-200 MBq (99m)Tc-HMPAO leukocytes. The leukocyte labelling efficiency was 65.5% and 89.0%, respectively, for the LPP and HBSS methods. There were 22 true positive, seven true negative and one false negative result in the LPP group, while in the HBSS group results were 18, nine and one, respectively. Diagnostic accuracy was similar with both methods although sensitivity was slightly higher in the HBSS group. In summary, these date indicate that leukocyte scintigraphy labelling using HBSS as the resuspension medium should be used as a first option for white blood cell labelling and diagnosis of inflammatory bowel disease.