RESUMO
BACKGROUND AND PURPOSE: Systemic lupus erythematosus (SLE) is an immune-mediated disease that may affect the nervous system. We explored the topographical organization of structural and functional brain connectivity in patients with SLE and its correlation with neuropsychiatric (NP) involvement and autoantibody profiles. METHODS: Graph theoretical analysis was applied to diffusion tensor magnetic resonance imaging (MRI) and resting-state functional MRI data from 32 patients with SLE and 32 age- and sex-matched healthy controls. Structural and functional connectivity matrices between 116 cortical/subcortical brain regions were estimated using a bivariate correlation analysis, and global and nodal network metrics were calculated. RESULTS: Structural, but not functional, global network properties (strength, transitivity, global efficiency and path length) were abnormal in patients with SLE versus controls (P < 0.0001), especially in patients with anti-double-stranded DNA (ADNA) autoantibodies (P = 0.03). No difference was found according to NP involvement or anti-phospholipid autoantibody status. Patients with SLE and controls shared identical structural hubs and the majority of functional hubs. In patients with SLE, all structural hubs showed reduced strength and clustering coefficient compared with controls (P from 0.001 to <0.0001), especially in patients with ADNA autoantibodies. Only a few differences in functional hub properties were found between patients with SLE and controls. Structural and functional hub measures did not differ according to NP involvement or anti-phospholipid autoantibody status. Significant correlations were found between clinical, MRI and network measures (r from -0.56 to 0.60, P from 0.0003 to 0.05). CONCLUSIONS: Abnormalities of global and nodal structural connectivity occur in patients with SLE, especially with ADNA autoantibodies, with a diffuse disruption of structural integrity. Functional network integrity may contribute to preserve clinical functions.
Assuntos
Encéfalo/patologia , Conectoma , Lúpus Eritematoso Sistêmico/patologia , Adulto , Anticorpos Antifosfolipídeos/análise , Anticorpos Antifosfolipídeos/imunologia , Autoanticorpos/imunologia , Encéfalo/diagnóstico por imagem , Córtex Cerebral/patologia , Análise por Conglomerados , DNA/imunologia , Imagem de Tensor de Difusão , Feminino , Humanos , Lúpus Eritematoso Sistêmico/diagnóstico por imagem , Lúpus Eritematoso Sistêmico/imunologia , Imageamento por Ressonância Magnética , Masculino , Pessoa de Meia-Idade , Adulto JovemRESUMO
BACKGROUND: In patients with moderate and severe paediatric traumatic brain injury (TBI), we investigated the presence and severity of white matter (WM) tract damage, cortical lobar and deep grey matter (GM) atrophies, their interplay and their correlation with outcome rating scales. METHODS: Diffusion tensor (DT) and 3D T1-weighted MRI scans were obtained from 22 TBI children (13 boys; mean age at insult = 11.6 years; 72.7% in chronic condition) and 31 age-matched healthy children. Patients were tested with outcome rating scales and the Wechsler Intelligence Scale for Children (WISC). DT MRI indices were obtained from several supra- and infra-tentorial WM tracts. Cortical lobar and deep GM volumes were derived. Comparisons between patients and controls, and between patients in acute (<6 months from the event) vs. chronic (≥6 months) condition were performed. RESULTS: Patients showed a widespread pattern of decreased WM FA and GM atrophy. Compared to acute, chronic patients showed severer atrophy in the right frontal lobe and reduced FA in the left inferior longitudinal fasciculus and corpus callosum (CC). Decreased axial diffusivity was observed in acute patients versus controls in the inferior fronto-occipital fasciculus and CC. Chronic patients showed increased axial diffusivity in the same structures. Uncinate fasciculus DT MRI abnormalities correlated with atrophy in the frontal and temporal lobes. Hippocampal atrophy correlated with reduced WISC scores, whereas putamen atrophy correlated with lower functional independence measure scores. CONCLUSIONS: The study isolated a distributed fronto-temporal network of structures particularly vulnerable to axonal damage and atrophy that may contribute to cognitive deficits following TBI.
Assuntos
Lesões Encefálicas Traumáticas/patologia , Substância Cinzenta/patologia , Rede Nervosa/patologia , Substância Branca/patologia , Adolescente , Atrofia/patologia , Lesões Encefálicas Traumáticas/diagnóstico por imagem , Criança , Imagem de Tensor de Difusão , Feminino , Substância Cinzenta/diagnóstico por imagem , Humanos , Imageamento por Ressonância Magnética , Masculino , Rede Nervosa/diagnóstico por imagem , Índice de Gravidade de Doença , Substância Branca/diagnóstico por imagemRESUMO
In this study, we describe the results of virological investigations carried out on cases of gastroenteritis reported in different communities within a 2-year pilot surveillance programme (January 2012 to December 2013) in the autonomous province of Bolzano (Northern Italy). Among the 162 norovirus (NoV)-positive cases out of 702 cases investigated, 76 were grouped in nine suspected outbreaks, 37 were hospital-acquired and 49 were community-acquired sporadic cases. NoV infections were found in all age groups in outbreak and community-acquired cases, while the highest peak of hospital-acquired infections occurred in the elderly. Sequence analyses helped to identify suspected outbreaks both in the community and in hospital wards. Although GII.4 is the predominant genotype, sequence data confirmed that at least seven genotypes circulate causing sporadic cases. Findings in this study confirmed the relevance of NoV infections as a cause of outbreaks, and impact of NoV infections in community-acquired sporadic cases in adults that are rarely described because of a lack of reporting.
Assuntos
Infecções por Caliciviridae/epidemiologia , Surtos de Doenças , Gastroenterite/epidemiologia , Norovirus/isolamento & purificação , Adolescente , Adulto , Idoso , Infecções por Caliciviridae/virologia , Criança , Pré-Escolar , Infecção Hospitalar/epidemiologia , Infecção Hospitalar/virologia , Monitoramento Epidemiológico , Feminino , Gastroenterite/virologia , Genótipo , Humanos , Lactente , Itália/epidemiologia , Masculino , Pessoa de Meia-Idade , Norovirus/genética , Fases de Leitura Aberta , Filogenia , Projetos Piloto , RNA Viral/análise , Análise de Sequência de RNA , Adulto JovemRESUMO
OBJECTIVES: Active myelination during childhood may influence the impact of multiple sclerosis (MS) on brain structural integrity. We studied normal-appearing white matter (NAWM) in children with MS onset before age 12 years using diffusion tensor (DT) magnetic resonance imaging (MRI). METHODS: DT MRI scans were obtained from 22 MS children with their first attack before age 12 years, and 31 healthy controls from two referral centers. Using probabilistic tractography, brain tissue integrity within interhemispheric, intrahemispheric, and projection tracts was compared between patients and site-matched controls. The impact of disease and age at MRI on tract NAWM fractional anisotropy (FA) and mean diffusivity (MD) values was evaluated using linear models. RESULTS: Compared to controls, pediatric MS patients had reduced FA and increased MD of the bilateral superior longitudinal fasciculus and corpus callosum (CC), without center-by-group interaction. CC NAWM average FA was correlated with brain T2 lesion volume. In controls, the majority of the tracts analyzed showed a significant increase of FA and decrease of MD with age. Such a linear correlation was lost in patients. CONCLUSIONS: In very young pediatric MS patients, DT MRI abnormalities affect brain WM tracts differentially, and are only partially correlated with focal WM lesions. Impaired maturation of WM tracts with age may be an additional factor contributing to these findings.
Assuntos
Imagem de Tensor de Difusão , Esclerose Múltipla/diagnóstico por imagem , Adolescente , Idade de Início , Anisotropia , Criança , Pré-Escolar , Corpo Caloso/diagnóstico por imagem , Imagem de Difusão por Ressonância Magnética/métodos , Imagem de Tensor de Difusão/métodos , Feminino , Humanos , Modelos Lineares , Masculino , Substância Branca/diagnóstico por imagemRESUMO
OBJECTIVES: Involvement of selected central nervous system (CNS) regions has been associated with depression and fatigue in MS. We assessed whether specific regional patterns of lesion distribution and atrophy of the gray (GM) and white matter (WM) are associated with these symptoms in MS. METHODS: Brain dual-echo and 3D T1-weighted images were acquired from 123 MS patients (69 depressed (D), 54 non-depressed (nD), 64 fatigued, 59 non-fatigued) and 90 controls. Lesion distribution, GM and WM atrophy were estimated using VBM and SPM8. RESULTS: Gender, age, disease duration and conventional MRI characteristics did not differ between D-MS and nD-MS patients. Fatigued patients experienced higher EDSS and depression than non-fatigued ones. Lesion distribution and WM atrophy were not related to depression and fatigue. Atrophy of regions in the frontal, parietal and occipital lobes had a combined effect on depression and fatigue. Atrophy of the left middle frontal gyrus and right inferior frontal gyrus were selectively related to depression. No specific pattern of GM atrophy was found to be related to fatigue. CONCLUSIONS: Depression in MS is linked to atrophy of cortical regions located in the bilateral frontal lobes. A distributed pattern of GM atrophy contributes to the concomitant presence of depression and fatigue in these patients.
Assuntos
Encéfalo/patologia , Depressão/patologia , Fadiga/etiologia , Fadiga/patologia , Esclerose Múltipla/patologia , Adulto , Depressão/etiologia , Feminino , Humanos , Interpretação de Imagem Assistida por Computador , Imageamento por Ressonância Magnética , Masculino , Pessoa de Meia-Idade , Esclerose Múltipla/complicaçõesRESUMO
OBJECTIVE: Using diffusion tensor magnetic resonance imaging (DT MRI), we analyzed the architectural integrity of the brain white matter (WM) from a large cohort of MS patients to identify the structural substrates of the concomitant presence of depression and fatigue. METHODS: Brain dual-echo, 3D T1-weighted and DT MRI scans were acquired from 147 MS patients and 90 gender- and age-matched healthy controls (HCs). Patients were stratified by the presence of depression (92 depressed (D), 55 not depressed (nD)) and fatigue (81 fatigued (F), 66 not fatigued (nF)). Sixty-five patients had co-occurrence of depression and fatigue (DF). Whole-brain voxel-wise comparisons of WM DT MRI abnormalities were performed using tract-based-spatial-statistics (TBSS). Tract-specific analyses were run in brain WM tracts using standard-space templates. RESULTS: Whole-brain voxel-wise analysis yielded no significant differences between patient subgroups. At tract-specific analysis, DF patients had reduced fractional anisotropy (FA) of the forceps minor. Reduced FA of the right anterior thalamic radiation and right uncinate fasciculus was found in F-MS vs not F-MS patients after correcting for depression. No significant differences were found between D vs not D-MS patients, after correcting for fatigue. CONCLUSIONS: This study provides evidence for partially overlapping damage to frontal and fronto-temporal pathways underlying depression and fatigue in MS.
Assuntos
Depressão/patologia , Fadiga/patologia , Esclerose Múltipla/patologia , Adulto , Idoso , Depressão/complicações , Transtorno Depressivo/complicações , Transtorno Depressivo/patologia , Imagem de Tensor de Difusão/métodos , Fadiga/complicações , Feminino , Humanos , Imageamento por Ressonância Magnética/métodos , Masculino , Pessoa de Meia-Idade , Esclerose Múltipla/complicações , Fibras Nervosas Mielinizadas/patologia , Rede Nervosa/patologiaRESUMO
BACKGROUND: Pathologic and magnetic resonance imaging (MRI) studies have shown that cortical lesions (CLs) are a frequent finding in multiple sclerosis (MS). OBJECTIVE: To quantify microstructural damage in CLs and normal appearing (NA) cortex in relapse-onset MS patients at different stages of the disease. METHODS: Brain double inversion recovery (DIR), diffusion tensor (DT) MRI and 3D T 1-weighted scans were acquired from 35 relapsing-remitting (RR) patients, 23 secondary progressive (SP) patients, 12 benign (B) MS patients and 41 healthy controls (HC). Diffusivity values in CLs, cortex, white matter (WM) lesions and normal-appearing white matter (NAWM) were assessed. RESULTS: Compared to HC, MS patients had a significantly lower fractional anisotropy (FA) and higher mean diffusivity (MD) in the cortex and NAWM. CLs had higher FA vs HC cortex and vs patients' cortex. Compared to RRMS patients, SPMS patients had higher WM lesion volume, higher MD in the cortex, and more severe damage to the NAWM and WM lesions. Compared to SPMS patients, BMS patients had lower MD and FA of CLs. Damage in other compartments was similar between SPMS and BMS patients. Damage in CLs had a high power to discriminate BMS from SPMS (area under the curve: 79-91%), with high specificity (85%), sensitivity (100%) and accuracy (90%). CONCLUSIONS: Microstructural imaging features of CLs differ from those of WM lesions and are likely to reflect neuronal damage and microglial activation. The nature and extent of CL damage can be used to help distinguish the different MS clinical phenotypes.
Assuntos
Encéfalo/patologia , Esclerose Múltipla/patologia , Adulto , Idoso , Anisotropia , Feminino , Humanos , Interpretação de Imagem Assistida por Computador , Imageamento por Ressonância Magnética/métodos , Masculino , Pessoa de Meia-Idade , Fibras Nervosas Mielinizadas/patologia , Adulto JovemRESUMO
AIM: We compared Hepatitis C virus (HCV) genotyping by direct sequencing of the non-structural 5b region (NS5b) and a commercial PCR/hybridization method based on the conserved 5´-untranslated region (5'UTR). METHODS: One hundred twenty HCV containing plasma samples were analyzed by NS5b sequencing with focus on samples with undetermined results or 1b subtype identification in the used combination of Cobas® AmpliPrep/Cobas® TaqMan96® PCR and subsequent Versant® HCV Genotype 2.0 Assay (LiPA). RESULTS: There was 100% concordance between the two methods for genotyping but only 83% for subtyping. Seventeen samples were designated 1b by hybridization but subtype 1a by NS5b sequencing. This is a general 5'UTR problem as the discordant results were additionally confirmed by 5'UTR sequencing. Thus our routine combination not only misclassified 38.6% of subtype 1a isolates as 1b but in contrast to NS5b sequencing was unable to discriminate between subtypes 2a/c, or 4a/c/d and also failed on a newly described subtype (10a/3k). [corrected]. CONCLUSIONS: [corrected] The applied 5'UTR methods allow the rapid determination of HCV genotypes but failed to correctly identify the subtype in many samples. This has implications for epidemiological studies or forensic evaluation of chains of infection and NS5b sequencing therefore is our method of choice under those circumstances.
Assuntos
Regiões 5' não Traduzidas/genética , Técnicas de Genotipagem/métodos , Hepacivirus/genética , Hibridização Genômica Comparativa/métodos , Genótipo , Humanos , Reação em Cadeia da Polimerase/métodos , Análise de Sequência de DNA/métodosRESUMO
Using diffusion tensor (DT) tractography, we quantified optic radiation (OR) structural changes in seven migraine patients with (MA) and eight without visual aura (MoA) and their relation to clinical manifestations and T2-visible burden. The corticospinal tract and the corpus callosum were studied as 'control' white matter (WM). No difference was found for any of the WM fibre bundles metrics between controls and MoA patients. MA patients had reduced average fractional anisotropy (FA) of both OR compared with controls and reduced average FA of the right OR compared with MoA patients. They also showed higher right OR mean diffusivity than controls. OR metrics were not correlated with clinical and magnetic resonance imaging (MRI) metrics. DT tractography reveals OR changes in MA patients that might represent a phenotypic biomarker of the disease given the lack of correlation with clinical and structural MRI metrics.
Assuntos
Imagem de Difusão por Ressonância Magnética , Transtornos de Enxaqueca/patologia , Transtornos de Enxaqueca/fisiopatologia , Vias Visuais/patologia , Vias Visuais/fisiopatologia , Adulto , Anisotropia , Atrofia , Corpo Caloso/citologia , Corpo Caloso/fisiologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Fibras Nervosas Mielinizadas/patologia , Tratos Piramidais/citologia , Tratos Piramidais/fisiologiaRESUMO
SUMMARY: Diffusion tensor (DT) MR imaging has frequently been applied in multiple sclerosis (MS) because of its ability to detect and quantify disease-related changes of the tissue microstructure within and outside T2-visible lesions. DT MR imaging data collection places high demands on scanner hardware and, though the acquisition and postprocessing can be relatively straightforward, numerous challenges remain in improving the reproducibility of this technique. Although there are some issues concerning image quality, echo-planar imaging is the most widely used acquisition scheme for diffusion imaging studies. Once the DT is estimated, indexes conveying the size, shape, and orientation of the DT can be calculated and further analyzed by using either histogram- or region-of-interest-based analyses. Because the orientation of the DT reflects the orientation of the axonal fibers of the brain, the pathways of the major white matter tracts can also be visualized. The DT model of diffusion, however, is not sufficient to characterize the diffusion properties of the brain when complex populations of fibers are present in a single voxel, and new ways to address this issue have been proposed. Two developments have enabled considerable improvements in the application of DT MR imaging: high magnetic field strengths and multicoil receiver arrays with parallel imaging. This review critically discusses models, acquisition, and postprocessing approaches that are currently available for DT MR imaging, as well as their limitations and possible improvements, to provide a better understanding of the strengths and weaknesses of this technique and a background for designing diffusion studies in MS.
Assuntos
Encéfalo/patologia , Imagem de Difusão por Ressonância Magnética/métodos , Esclerose Múltipla/patologia , HumanosRESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: It is not always possible to correlate the "emic" terms to the "etic" ones during ethnopharmacological surveys, especially regarding those related to Culture-Bound Syndromes (CBS). Nevertheless, it is the role of ethnopharmacology to address these correlations, since they are the basis for the understanding of potential bioactives. AIM AND OBJECTIVES: This study reports the clinical manifestations and therapeutic resources used for the treatment of CBS among some riverine inhabitants of Brazilian Amazonia. An effort was made to establish a correspondence between the local "emic" terms of traditional medicine and the symptoms or diseases known by conventional medicine ("etic" terms). The ultimate goal was to gain insights to suggest further pharmacological studies with the local resources. MATERIAL AND METHODS: Fieldwork was guided by methods of anthropology, botany and zoology-with the assistance of a doctor-among the traditional healing experts in Jaú National Park (during 199 days in 1995) and Unini River Extractive Reserve (210 days from 2008 to 2012). RESULTS: Fifty-nine healers of different kinds were interviewed: a prayer-maker, medium, natural resource expert, massage therapist, midwife and snakebite healer. The clinical manifestations and healing resources of the following CBS were collected: "mau olhado" (evil eye), "quebrante" (chipping); "espante" (fright or susto); "doença do ar" (air diseases); "vento caído" (fallen wind); "derrame" (leakage); "mãe do corpo" (mother of the body) and "panema" (unlucky). The first three seem to be local variations of other CBSs already described in Latin America. "doença do ar", "vento caído", "derrame" and "mãe do corpo" seem to be folk terms for known conventional medical disorders, while "panema" is a yet undescribed Brazilian CBS that is possibly related to dysthymic disorder or depression and deserves further investigation. Treatments included prayer rituals, fumigation, baths and oral remedies using 25 plants and 10 animals. CONCLUSION: It was possible to establish hypothetical correlations between CBS as described by the riverine population studied and some "etic" terms. The main importance of this is to help the proposition of target-oriented pharmacological studies of the natural resources used by these communities. Accordingly, the following plants are suggested to be submitted to further studies for antidepressant and anxiolytic activities: Siparuna guianensis, Mansoa alliacea, Leucas martinicensis, Petiveria alliacea, Annona montana and Alpinia nutans; for anti-seizure activity: Protium amazonicum, Protium aracouchini and Protium heptaphyllum; finally for antispasmodic activity: Leucas martinicensis.
Assuntos
Medicina Tradicional/métodos , Preparações de Plantas/farmacologia , Plantas Medicinais/química , Animais , Brasil , Características Culturais , Etnofarmacologia , Feminino , Humanos , Entrevistas como Assunto , Masculino , Fitoterapia/métodosRESUMO
Ultrasonographic evaluation of the adrenal glands was performed in 85 dogs, followed by macroscopic and histopathological examination either post-mortem or after adrenalectomy. This retrospective cross-sectional study evaluated the difference between gross and ultrasonographic measurements to determine the diagnostic accuracy of ultrasonography in the evaluation of canine adrenal gland size. The differences were assessed for gland length, thickness at cranial, middle and caudal regions, and surface area. In our sample, ultrasound error accuracy ranged between 0% in measurement of the right adrenal gland surface area and 25.21% for left cranial pole thickness. The parameters with minor errors were caudal pole thickness (3.64% right side and 3.49% left side) and length (5.75% right side and 2.19% left side). The ultrasonographic measurements generally underestimated the actual size of the adrenal glands. No statistically significant differences were observed for measurement errors between normal and pathological adrenal glands. This study confirmed that the caudal pole of both glands is the best parameter for ultrasonographic evaluation of normal and pathological adrenal glands size in dog. Furthermore, the surface area could be considered as a dimensional parameter for better assessment of the complex shape and the global aspect of the adrenal glands, while standardize ultrasonographic projections are needed to measure the cranial pole of both adrenal glands.
Assuntos
Glândulas Suprarrenais/diagnóstico por imagem , Glândulas Suprarrenais/cirurgia , Erros de Diagnóstico/veterinária , Tamanho do Órgão/fisiologia , Ultrassonografia/veterinária , Glândulas Suprarrenais/anatomia & histologia , Animais , Estudos Transversais , Cães , Estudos RetrospectivosRESUMO
BACKGROUND: The pathophysiology of abdominal distention in irritable bowel syndrome (IBS) is still a matter of debate, but the relationship between modifications of intestinal tone and abdominal volume has never been analyzed. METHODS: Eighty-four patients affected by IBS and reporting moderate to severe abdominal distention were enrolled. Thirty-nine presented abdominal distention immediately after and forty-five presented abdominal distention independently of meal intake. Twenty healthy volunteers (HV), comparable for gender and age, were also enrolled. All the subjects underwent fasting and postprandial recto-sigmoid volume monitoring with barostat and abdominal girth measurement to evaluate abdominal distention. KEY RESULTS: In comparison with HV (75±13 mL) and with patients with meal-unrelated abdominal distention (135±56 mL), in the subgroup of patients with severe meal-related abdominal distention recto-sigmoid tone response to the meal was significantly reduced (mean increase of balloon volume 184±89 mL; P<.001), paralleling abdominal girth increase and occurring immediately after test meal intake. Meal-induced abdominal girth modification was significantly correlated with meal-related modification of recto-sigmoid tone (r=.71) and abdominal symptoms. CONCLUSIONS: In patients with IBS suffering from severe postprandial abdominal distention, a postprandial reduction of intestinal tone is associated with this bothersome symptom. Further studies are needed to evaluate whether drugs acting on the modification of intestinal tone could be useful in the treatment of these patients.
Assuntos
Síndrome do Intestino Irritável/fisiopatologia , Tono Muscular/fisiologia , Músculo Liso/fisiopatologia , Adulto , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Período Pós-Prandial , Adulto JovemRESUMO
BACKGROUND AND PURPOSE: The automatic segmentation of MS lesions could reduce time required for image processing together with inter- and intraoperator variability for research and clinical trials. A multicenter validation of a proposed semiautomatic method for hyperintense MS lesion segmentation on dual-echo MR imaging is presented. MATERIALS AND METHODS: The classification technique used is based on a region-growing approach starting from manual lesion identification by an expert observer with a final segmentation-refinement step. The method was validated in a cohort of 52 patients with relapsing-remitting MS, with dual-echo images acquired in 6 different European centers. RESULTS: We found a mathematic expression that made the optimization of the method independent of the need for a training dataset. The automatic segmentation was in good agreement with the manual segmentation (dice similarity coefficient = 0.62 and root mean square error = 2 mL). Assessment of the segmentation errors showed no significant differences in algorithm performance between the different MR scanner manufacturers (P > .05). CONCLUSIONS: The method proved to be robust, and no center-specific training of the algorithm was required, offering the possibility for application in a clinical setting. Adoption of the method should lead to improved reliability and less operator time required for image analysis in research and clinical trials in MS.
Assuntos
Cólera , Gastroenteropatias/microbiologia , Adulto , Pré-Escolar , Fezes/microbiologia , Feminino , Humanos , MasculinoRESUMO
The expression of telomerase activity and the in situ localization of the human telomerase RNA component (hTR) in melanocytic skin lesions was evaluated in specimens from sixty-three patients. Specimens of melanocytic nevi, primary melanomas and subcutaneous metastases of melanoma were obtained from fifty-eight patients, whereas metastasized lymph nodes were obtained from five patients. Telomerase activity was determined in these specimens by using a Polymerase Chain Reaction-based assay (TRAP). High relative mean telomerase activity levels were detected in metastatic melanoma (subcutaneous metastases = 54.5, lymph node metastases = 56.5). Much lower levels were detected in primary melanomas, which increased with advancing levels of tumor cell penetration (Clark II = 0.02, Clark III = 1.1, and Clark IV = 1.9). Twenty-six formalin-fixed, paraffin-embedded melanocytic lesions were sectioned and analyzed for telomerase RNA with a radioactive in situ hybridization assay. In situ hybridization studies with a probe to the template RNA component of telomerase confirmed that expression was almost exclusively confined to tumor cells and not infiltrating lymphocytes. These results indicate that levels of telomerase activity and telomerase RNA in melanocytic lesions correlate well with clinical stage and could potentially assist in the diagnosis of borderline lesions.
Assuntos
Melanoma/enzimologia , Nevo Pigmentado/enzimologia , Neoplasias Cutâneas/enzimologia , Telomerase/biossíntese , Humanos , Hibridização In Situ , Melanoma/patologia , Melanoma/secundário , Mitose , Invasividade Neoplásica , Nevo Pigmentado/patologia , RNA/análise , Telomerase/genéticaRESUMO
The vascular endothelial growth factor is produced by a large variety of human tumors, including melanoma, in which it appears to play an important role in the process of tumor-induced angiogenesis. Little information is available on the role of placenta growth factor, a member of the vascular endothelial growth factor family of cytokines, in tumor angiogenesis, even though placenta growth factor/vascular endothelial growth factor heterodimers have been recently isolated from tumor cells. To investigate the role of placenta growth factor and vascular endothelial growth factor homodimers and heterodimers in melanoma angiogenesis and growth, 19 human melanoma cell lines derived from primary or metastatic tumors were characterized for the expression of these cytokines and their receptors. Release of placenta growth factor and vascular endothelial growth factor polypeptides into the supernatant of human melanoma cells was demonstrated. Reverse transcriptase polymerase chain reaction analysis showed the presence of mRNAs encoding at least three different vascular endothelial growth factor isoforms (VEGF(121), VEGF(165), and VEGF(189)) and transcripts for two placenta growth factor isoforms (PlGF-1 and PlGF-2) in human melanoma cells. In addition, placenta growth factor expression in human melanoma in vivo was detected by immunohistochemical staining of tumor specimens. Both primary and metastatic melanoma cells were found to express the mRNAs encoding for vascular endothelial growth factor and placenta growth factor receptors (KDR, Flt-1, neuropilin-1, and neuropilin-2), and exposure of melanoma cells to these cytokines resulted in a specific proliferative response, supporting the hypothesis of a role of these angiogenic factors in melanoma growth. J Invest Dermatol 115:1000-1007 2000
Assuntos
Fatores de Crescimento Endotelial/farmacologia , Linfocinas/farmacologia , Melanoma/metabolismo , Melanoma/patologia , Dimerização , Humanos , Fator de Crescimento Placentário , Proteínas da Gravidez/farmacologia , Isoformas de Proteínas , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Neoplasias Cutâneas/metabolismo , Neoplasias Cutâneas/patologia , Fator A de Crescimento do Endotélio Vascular , Fatores de Crescimento do Endotélio VascularRESUMO
Two series of medorinone (3) analogs were prepared by modifications at C(2) and C(5). The C(2)-series was prepared from 2-chloro-5-methyl-1,6-naphthyridine (4) by replacement of the chloro group with various nucleophiles. The C(5)-series was prepared from 5-acyl-6-[2-(dimethylamino)-ethenyl]-2(1H)-pyridinone (11), 5-bromo-1,6-naphthyridin-2(1H)-one (17), and 1,3-diketones 19 and 27. 1,6-Naphthyridin-2(1H)-ones are novel inhibitors of cAMP PDE III. Modification of the carbonyl group of 3 or N-methylation at N(1) resulted in a dramatic loss of enzyme activity. Absence of the C(5)-methyl group of medorinone (3) or its shift to C(3) or C(7) also resulted in reduced activity. Substitution at C(3) also diminished activity. However, substitution at C(5) by a wide variety of substituents led to improvement of enzyme activity and several C(5)-substituted analogs were more potent than milrinone.
Assuntos
3',5'-AMP Cíclico Fosfodiesterases/antagonistas & inibidores , Inibidores Enzimáticos/síntese química , Naftiridinas/síntese química , Animais , Cardiotônicos/síntese química , Cardiotônicos/química , Cardiotônicos/farmacologia , Cães , Inibidores Enzimáticos/química , Inibidores Enzimáticos/farmacologia , Naftiridinas/química , Naftiridinas/farmacologia , Relação Estrutura-AtividadeRESUMO
The transformation of 3-bromo-1,6-naphthyridin-2(1H)-ones 8 to thiazolo[4,5-b][1,6]naphthyridin-2(1H)-ones 12 resulted in a 2-9-fold increase in cAMP phosphodiesterase (PDE) III inhibitory potency. Unlike the secondary binding sites on the cAMP PDE III isozyme which interact with the methyl group of milrinone (2) and CI-930 (4), the site which interacts with the 5-substituents of 1,6-naphthyridin-2(1H)-ones and the 8-substituents of thiazolo[4,5-b][1,6]naphthyridin-2(1H)-ones 12 is able to accommodate a diverse group of substituents which have different steric and electronic requirements.
Assuntos
3',5'-AMP Cíclico Fosfodiesterases/antagonistas & inibidores , Naftiridinas/farmacologia , Espectroscopia de Ressonância Magnética , Naftiridinas/químicaRESUMO
We have analysed the [AGC] expansion in leucocytes, muscle and sperm from 17 individuals affected by myotonic dystrophy (DM). Skeletal muscle showed a larger repeat number than leucocytes in the same patient. A similar degree of expansion was detected in differently affected muscles of a single patient. The germline mutation (< or = 350 repeats) was expanded in somatic cells of the progeny in all patients examined. Our results provide evidence of an early postzygotic instability of the [AGC] repeat in DM.