RESUMO
Skeletal muscle tissue differs with regard to the abundance of glycolytic and oxidative fiber types. In this context, capillary density has been described to be higher in muscle tissue with more oxidative metabolism as compared to that one with more glycolytic metabolism, and the highest abundance of capillaries has been found in boneward-oriented moieties of skeletal muscle tissue. Importantly, capillary formation is often analyzed as a measure for angiogenesis, a process that describes neo-vessel formation emanating from preexisting vessels, occurring, i.e., after arterial occlusion. However, a standardized way for investigation of calf muscle capillarization after surgically induced unilateral hind limb ischemia in mice, especially considering these locoregional differences, has not been provided so far. In this manuscript, a novel, methodical approach for reliable analysis of capillary density was established using anatomic-morphological reference points, and a software-assisted way of capillary density analysis is described. Thus, the systematic approach provided conscientiously considers intra-layer differences in capillary formation and therefore guarantees for a robust, standardized analysis of capillary density as a measure for angiogenesis. The significance of the methodology is further supported by the observation that capillary density in the calf muscle layers analyzed negatively correlates with distal lower limb perfusion measured in vivo.
Assuntos
Capilares/crescimento & desenvolvimento , Membro Posterior/irrigação sanguínea , Músculo Esquelético/irrigação sanguínea , Neovascularização Fisiológica , Animais , Modelos Animais de Doenças , Membro Posterior/patologia , Imuno-Histoquímica , Isquemia/patologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Microscopia de Fluorescência , Músculo Esquelético/patologiaRESUMO
BACKGROUND AND PURPOSE: Worldwide, oesophageal cancer is the eighth most common cancer and has a very poor survival rate. In order to identify new tolerable treatment options for oesophageal squamous cell carcinoma (ESCC), erlotinib was tested with moderate efficacy in phase I and II studies. As 4-methylumbelliferone (4-MU), an hyaluronan (HA) synthesis inhibitor showed anti-cancer effects in vitro, and in ESCC xenograft tumours, we investigated whether the anti-cancer effects of erlotinib could be augmented by combining it with 4-MU. EXPERIMENTAL APPROACH: ESCC cell lines were treated with erlotinib or gefitinib (1 µmol·L-1 ) and 4-MU (300 µmol·L-1 ), and the cell count, cell cycle progression and migration were determined as compared to the single agents and the solvent-control. KEY RESULTS: The combination of erlotinib and 4-MU synergistically inhibited the proliferation of ESCC cell lines. Furthermore, the migration speed of ESCC cell line KYSE-410 in gap closure assays was significantly reduced by the combination of erlotinib and 4-MU. Decreased ERK phosphorylation could explain the anti-proliferative and anti-migratory effects in the combined treatment group. Finally, the combination was additionally able to decrease the growth of multicellular tumour spheroids, a three-dimensional cell culture model that was associated with sustained inhibition of ERK1/2 phosphorylation. CONCLUSIONS AND IMPLICATIONS: The combination of 4-MU and erlotinib showed promising anti-cancer efficacies in the ESCC cell lines.
RESUMO
BACKGROUND AND PURPOSE: Combined hormone replacement therapy with oestrogens plus the synthetic progestin medroxyprogesterone acetate (MPA) is associated with an increased risk of thrombosis. However, the mechanisms of this pro-thrombotic effect are largely unknown. The purpose of this study was to: (i) compare the pro-thrombotic effect of MPA with another synthetic progestin, norethisterone acetate (NET-A), (ii) determine if MPA's pro-thrombotic effect can be antagonized by the progesterone and glucocorticoid receptor antagonist mifepristone and (iii) elucidate underlying mechanisms by comparing aortic gene expression after chronic MPA with that after NET-A treatment. EXPERIMENTAL APPROACH: Female apolipoprotein E-deficient mice were ovariectomized and treated with placebo, MPA, a combination of MPA + mifepristone or NET-A for 90 days on a Western-type diet. Arterial thrombosis was measured in vivo in a photothrombosis model. Aortic gene expression was analysed using microarrays; GeneOntology and KEGG pathway analyses were conducted. KEY RESULTS: MPA's pro-thrombotic effects were prevented by mifepristone, while NET-A did not affect arterial thrombosis. Aortic gene expression analysis showed, for the first time, that gestagens induce similar effects on a set of genes potentially promoting thrombosis. However, in NET-A-treated mice other genes with potentially anti-thrombotic effects were also affected, which might counterbalance the effects of the pro-thrombotic genes. CONCLUSIONS AND IMPLICATIONS: The pro-thrombotic effects of synthetic progestins appear to be compound-specific, rather than representing a class effect of gestagens. Furthermore, the different thrombotic responses elicited by MPA and NET-A might be attributed to a more balanced, 'homeostatic' gene expression induced in NET-A- as compared with MPA-treated mice.
Assuntos
Aorta/efeitos dos fármacos , Trombose das Artérias Carótidas/genética , Anticoncepcionais Femininos/farmacologia , Regulação da Expressão Gênica/efeitos dos fármacos , Acetato de Medroxiprogesterona/farmacologia , Noretindrona/análogos & derivados , Animais , Aorta/metabolismo , Apolipoproteínas E/deficiência , Apolipoproteínas E/genética , Células Cultivadas , Vasos Coronários/citologia , Células Endoteliais/efeitos dos fármacos , Células Endoteliais/metabolismo , Feminino , Perfilação da Expressão Gênica , Humanos , Camundongos Knockout , Miócitos de Músculo Liso/efeitos dos fármacos , Miócitos de Músculo Liso/metabolismo , Noretindrona/farmacologia , Acetato de Noretindrona , Análise de Sequência com Séries de Oligonucleotídeos , Ovariectomia , Reação em Cadeia da PolimeraseAssuntos
Angiografia Cerebral/métodos , Infarto da Artéria Cerebral Média/diagnóstico , Infarto da Artéria Cerebral Média/etiologia , Hemorragia Subaracnoídea Traumática/complicações , Hemorragia Subaracnoídea Traumática/diagnóstico , Vasoespasmo Intracraniano/diagnóstico , Vasoespasmo Intracraniano/etiologia , Adolescente , Diagnóstico Diferencial , Humanos , MasculinoRESUMO
OBJECTIVES: Thrombolysis in stroke remains underutilized in daily practice. We analyzed the impact of a multimodal strategy on the rate of thrombolysis and specific procedure times during the implementation of a community hospital stroke unit. MATERIAL AND METHODS: During a period of 2 years before and after implementation of a stroke unit, we prospectively recorded all patients with thrombolysis and specific procedure times. Calculated door-to-needle time (DNT), door-to-CT time (DCT) and CT-to-needle time (CNT) were analyzed. All structural changes before and after the implementation were analyzed. RESULTS: The number of patients with thrombolysis increased from 24 in 2005-2006 (4.8% of all admitted patients with ischemic stroke) to 95 in 2007-2008 (12.8%). DNT was significantly reduced from 62.2±36.1 to 38.5±22.2 min (P<0.001). DCT remained unchanged at 10.3±9.5 to 10.4±13.9 min (P=0.974), whereas CNT improved from 45.7±23.1 to 28.3±20.3 min (P=0.001). Several structural changes concerning staff, logistics, procedures and laboratory were identified which contributed to decreasing DNT. CONCLUSIONS: A multimodal strategy including several structural changes enables the successful implementation of a community hospital stroke unit offering rapid access to thrombolysis with a very short DNT.
Assuntos
Terapia Combinada/métodos , Serviços Médicos de Emergência/métodos , Hospitais Comunitários/métodos , Acidente Vascular Cerebral/tratamento farmacológico , Terapia Trombolítica/métodos , Adolescente , Adulto , Fatores Etários , Idoso , Idoso de 80 Anos ou mais , Hemorragia Cerebral/etiologia , Hemorragia Cerebral/mortalidade , Hemorragia Cerebral/prevenção & controle , Estudos de Coortes , Terapia Combinada/estatística & dados numéricos , Serviços Médicos de Emergência/estatística & dados numéricos , Serviço Hospitalar de Emergência , Feminino , Fibrinolíticos/uso terapêutico , Alemanha , Pesquisas sobre Atenção à Saúde/métodos , Hospitais Comunitários/estatística & dados numéricos , Humanos , Masculino , Medicina/estatística & dados numéricos , Medicina/tendências , Pessoa de Meia-Idade , Neurologia/estatística & dados numéricos , Neurologia/tendências , Equipe de Assistência ao Paciente/normas , Equipe de Assistência ao Paciente/estatística & dados numéricos , Equipe de Assistência ao Paciente/tendências , Estudos Prospectivos , Garantia da Qualidade dos Cuidados de Saúde/métodos , Qualidade da Assistência à Saúde/estatística & dados numéricos , Acidente Vascular Cerebral/diagnóstico , Acidente Vascular Cerebral/mortalidade , Terapia Trombolítica/estatística & dados numéricos , Fatores de Tempo , Ativador de Plasminogênio Tecidual/uso terapêutico , Tomografia Computadorizada por Raios X/estatística & dados numéricos , Transporte de Pacientes , Resultado do Tratamento , Adulto JovemRESUMO
BACKGROUND: After myocardial infarction (MI), extensive remodeling of extracellular matrix contributes to scar formation and preservation of hemodynamic function. On the other hand, adverse and excessive extracellular matrix remodeling leads to fibrosis and impaired function. The present study investigates the role of the small leucine-rich proteoglycan biglycan during cardiac extracellular matrix remodeling and cardiac hemodynamics after MI. METHODS AND RESULTS: Experimental MI was induced in wild-type (WT) and bgn(-/0) mice by permanent ligation of the left anterior descending coronary artery. Biglycan expression was strongly increased at 3, 7, and 14 days after MI in WT mice. bgn(-/0) mice showed increased mortality rates after MI as a result of frequent left ventricular (LV) ruptures. Furthermore, tensile strength of the LV derived from bgn(-/0) mice 21 days after MI was reduced as measured ex vivo. Collagen matrix organization was severely impaired in bgn(-/0) mice, as shown by birefringence analysis of Sirius red staining and electron microscopy of collagen fibrils. At 21 days after MI, LV hemodynamic parameters were assessed by pressure-volume measurements in vivo to obtain LV end-diastolic pressure, end-diastolic volume, and end-systolic volume. bgn(-/0) mice were characterized by aggravated LV dilation evidenced by increased LV end-diastolic volume (bgn(-/0), 111+/-4.2 microL versus WT, 96+/-4.4 microL; P<0.05) and LV end-diastolic pressure (bgn(-/0), 24+/-2.7 versus WT, 18+/-1.8 mm Hg; P<0.05) and severely impaired LV function (EF, bgn(-/0), 12+/-2% versus WT, 21+/-4%; P<0.05) 21 days after MI. CONCLUSIONS: Biglycan is required for stable collagen matrix formation of infarct scars and for preservation of cardiac hemodynamic function.
Assuntos
Proteínas da Matriz Extracelular/metabolismo , Infarto do Miocárdio/metabolismo , Proteoglicanas/metabolismo , Remodelação Ventricular/fisiologia , Análise de Variância , Animais , Biglicano , Cicatriz , Proteínas da Matriz Extracelular/deficiência , Proteínas da Matriz Extracelular/genética , Genótipo , Ruptura Cardíaca Pós-Infarto/metabolismo , Hemodinâmica , Humanos , Estimativa de Kaplan-Meier , Camundongos , Camundongos Knockout , Infarto do Miocárdio/mortalidade , Infarto do Miocárdio/patologia , Infarto do Miocárdio/fisiopatologia , Miocárdio/metabolismo , Miocárdio/patologia , Fenótipo , Proteoglicanas/deficiência , Proteoglicanas/genética , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
The aim of this prospective study was to determine the prevalence and characteristics of acid gastro-oesophageal reflux (GER) in patients with idiopathic pulmonary fibrosis (IPF). Sixty-five consecutive patients with well-defined IPF were subjected to 24-h pH monitoring and oesophageal manometry. A total of 133 consecutive patients with intractable asthma and symptoms of GER were used as comparisons. The prevalence of abnormal acid GER in IPF patients was 87%, with 76% and 63% demonstrating abnormal distal and proximal oesophageal acid exposures, respectively. Abnormal acid GER was significantly more common in IPF patients than asthma patients. Only 47% of IPF patients experienced classic GER symptoms. Despite treatment with standard doses of proton pump inhibitors (PPIs), 12 out of 19 patients receiving PPIs during the 24-h pH monitoring had abnormal oesophageal acid exposures by pH probe. There was no correlation between IPF severity and acid GER severity. In conclusion, abnormal acid gastro-oesophageal reflux is highly prevalent, but often clinically occult in patients with idiopathic pulmonary fibrosis. Standard doses of proton pump inhibitors may not suppress the acid gastro-oesophageal reflux in this population. Therefore, further studies are needed to determine if acid abnormal gastro-oesophageal reflux represents an important risk factor for idiopathic pulmonary fibrosis development or progression, and if optimal suppression of acid gastro-oesophageal reflux slows the progression of idiopathic pulmonary fibrosis and/or decreases episodic exacerbations of idiopathic pulmonary fibrosis.
Assuntos
Refluxo Gastroesofágico/etiologia , Fibrose Pulmonar/complicações , Adulto , Idoso , Idoso de 80 Anos ou mais , Distribuição de Qui-Quadrado , Feminino , Determinação da Acidez Gástrica , Refluxo Gastroesofágico/tratamento farmacológico , Refluxo Gastroesofágico/epidemiologia , Refluxo Gastroesofágico/fisiopatologia , Humanos , Concentração de Íons de Hidrogênio , Modelos Lineares , Masculino , Pessoa de Meia-Idade , Monitorização Ambulatorial , Prevalência , Estudos Prospectivos , Inibidores da Bomba de Prótons , Testes de Função Respiratória , Estatísticas não ParamétricasRESUMO
In a young patient with spinal angioma, the diagnosis was not established until the fourth episode of bleeding. Exemplified by this case, the clinical symptomatology of spinal subarachnoid hemorrhage is described, and the diagnostic and therapeutic procedures are reviewed. In patients with SAH it is necessary to look for spinal symptoms, especially if cerebral angiography proves negative. In diagnosing a spinal SAH priority should be given to myelography and/or MRT, followed by spinal angiography.
Assuntos
Hemangioma/diagnóstico , Neoplasias da Medula Espinal/diagnóstico , Hemorragia Subaracnóidea/diagnóstico , Adulto , Angiografia , Diagnóstico Diferencial , Embolização Terapêutica , Hemangioma/terapia , Humanos , Masculino , Recidiva , Medula Espinal/irrigação sanguínea , Neoplasias da Medula Espinal/terapia , Hemorragia Subaracnóidea/terapiaRESUMO
Renal medullary cells use sorbitol, betaine, and other organic compounds as osmotic effectors (osmolytes) to balance high extracellular NaCl. Excess sorbitol is also implicated in diabetes complications in several organs including kidneys. To study regulation of renal sorbitol, male Wistar rats were given an aldose reductase inhibitor, sorbinil, at 40 mg.kg-1.day-1 in food to block sorbitol formation from glucose. Inner medullas of kidneys were analyzed for osmolytes by high-performance liquid chromatography and atomic absorption. Animals on sorbinil had significantly reduced medullary sorbitol contents in a group on ad libitum water for 10 days (2.7 mmol/kg wet wt compared with 4.8 in controls) and in an antidiuretic group kept 7 days and an additional 3 days without water (3.8 mmol/kg wet wt compared with 7.2 in antidiuretic controls). In both groups, betaine contents were significantly elevated (9.2 mmol/kg wet wt compared with 5.5 in ad libitum water controls: 6.4 mmol/kg wet wt compared with 4.2 in antidiuretic controls). No other osmolytes differed. Total contents of nonurea organic osmolytes maintained a constant ratio to sodium contents; thus increased betaine concomitant with decreased sorbitol may have maintained constant cell volume. In contrast, in animals kept 21 days on sorbinil, there were significant decreases in urea and inositol contents. However, there were no significant differences in sorbitol or betaine compared with controls, suggesting a compensating increase in sorbitol production or in sorbinil removal.
Assuntos
Aldeído Redutase/antagonistas & inibidores , Imidazóis/farmacologia , Imidazolidinas , Medula Renal/metabolismo , Animais , Betaína/metabolismo , Linhagem Celular , Diuréticos/antagonistas & inibidores , Medula Renal/citologia , Masculino , Osmose , Ratos , Ratos EndogâmicosRESUMO
Information from a case-control study of all patients with cutaneous malignant melanoma first diagnosed in Scotland in 1987 has been used to derive a personal risk-factor chart that can be used by both the medical profession and the general public. The relative risk of cutaneous melanoma is estimated from the four strongest risk factors identified by conditional logistic regression. These factors are total number of benign pigmented naevi above 2 mm diameter; freckling tendency; number of clinically atypical naevi (over 5 mm diameter and having an irregular edge, irregular pigmentation, or inflammation); and a history of severe sunburn at any time in life. Use of this risk-factor chart should enable preventive advice for and surveillance of those at greatest risk.
Assuntos
Melanoma/etiologia , Melanose/complicações , Nevo Pigmentado/complicações , Neoplasias Cutâneas/etiologia , Queimadura Solar/complicações , Clima Tropical , Raios Ultravioleta/efeitos adversos , Adolescente , Adulto , Idoso , Criança , Feminino , Humanos , Masculino , Programas de Rastreamento , Melanoma/epidemiologia , Melanoma/prevenção & controle , Pessoa de Meia-Idade , Análise de Regressão , Fatores de Risco , Escócia , Índice de Gravidade de Doença , Neoplasias Cutâneas/epidemiologia , Neoplasias Cutâneas/prevenção & controle , Queimadura Solar/prevenção & controleRESUMO
Chronic rhinosinusitis is an extremely common clinical problem of which the etiology is poorly understood. To understand the role of common environmental antigens in this disease, natural immunity to antigens derived from the house dust mite was evaluated in 22 adults with chronic rhinosinusitis and compared to a carefully matched group of patients with chronic asthma or to a group of normal individuals. Allergic reactivity to dust mites was very common in patients with chronic rhinosinusitis, with 68% exhibiting a positive immediate skin test reaction and 41% exhibiting elevated levels of mite-specific serum IgE; 72% of patients with rhinosinusitis also exhibited markedly elevated levels of mite-specific serum IgG, which were present in both mite-allergic and nonallergic patients. IgG titers were much higher in the group with rhinosinusitis than in patients with asthma, whereas allergic reactivity to dust mites was significantly higher in the patients with asthma. Mite-specific immunity was low or absent in the group of normal individuals. These findings demonstrate that natural immunity to dust mites is very common in patients with chronic rhinosinusitis and suggest that immunity to mites may be involved in this syndrome. Furthermore, the data indicate that there may be significant differences in the ability of patients with rhinosinusitis or asthma to produce mite-specific antibodies of the IgG class.