RESUMO
As a target for clinical anti-cancer treatment, epidermal growth factor receptor (EGFR) exhibits its over-expression on various tumour cells and is associated with the development of a variety of human cancers. Herein, we described the synthesis, antiproliferative activity assay and 4D-QSAR studies of thiadiazole derivatives bearing acrylamide moiety as EGFR inhibitors. Compared with Gefitinib, some of the target compounds have excellent antiproliferative activities against EGFR-expressed A431 cell line. The robust and reliable 4D-QSAR was constructed using comparative distribution detection algorithm, ordered predictors selection and genetic algorithm method, and the following acceptable statistics are shown: r2 = 0.82, Q2LOO = 0.67, Q2LMO = 0.61, r2Pred = 0.78.
Assuntos
Antineoplásicos , Receptores ErbB , Relação Quantitativa Estrutura-Atividade , Humanos , Acrilamida , Antineoplásicos/farmacologia , Linhagem Celular Tumoral , Receptores ErbB/antagonistas & inibidoresRESUMO
Cannabinoid receptor has been shown to be overexpressed in various types of cancers, especially non-small cell lung cancer. As a result, it could be used as novel target for anticancer treatments. Because receptor-dependent 4D-QSAR generates conformational ensemble profiles of compounds by molecular dynamics simulations at the binding site of the enzyme, this work describes the synthesis, biological activity evaluation and 4D-QSAR studies of 4,5-dihydro-1,3,4-oxadiazole derivatives targeting cannabinoid receptor. Compared with WIN55,212-2, compound 5 f showed the best antiproliferative activity. The receptor-dependent 4D-QSAR model was generated by multiple linear regression method using QSARINS. Leave-n-out cross-validation and chemical applicability domain were performed to analyse the independent test set and to verify the robustness of the model. The best 4D-QSAR model showed the following statistics: r2 = 0.8487, Q2LOO = 0.7667, Q2LNO = 0.7524, and r2Pred = 0.8358.
Assuntos
Oxidiazóis/farmacologia , Relação Quantitativa Estrutura-Atividade , Receptores de Canabinoides/efeitos dos fármacos , Células A549 , Proliferação de Células/efeitos dos fármacos , Humanos , Conformação Molecular , Simulação de Dinâmica Molecular , Oxidiazóis/síntese química , Oxidiazóis/químicaRESUMO
OBJECTIVE: To compare outcomes between intensivist-directed and cardiac surgeon-directed care delivery models. DESIGN: This retrospective, historical-control study was performed in a cohort of adult cardiac surgical patients at Zhongshan Hospital (Fudan University, China). During the first phase (March to August 2015), cardiac surgeons were in charge of postoperative care while intensivists were in charge during the second phase (September 2015-June 2016). Both phases were compared regarding successful extubation rate, intensive care unit (ICU) length of stay (LOS), and in-hospital mortality. SETTING: Tertiary Zhongshan Hospital (Fudan University, China). PATIENTS: Consecutive adult patients admitted to the cardiac surgical ICU (CSICU) after heart surgery. INTERVENTIONS: Phase I patients treated by cardiac surgeons, and phase II patients treated by intensivists. MAIN VARIABLES OF INTEREST: Successful extubation, ICU LOS and in-hospital mortality. RESULTS: A total of 1792 (phase I) and 3007 patients (phase II) were enrolled. Most variables did not differ significantly between the two phases. However, patients in phase II had a higher successful extubation rate (99.17% vs. 98.55%; p=0.043) and a shorter median duration of mechanical ventilation (MV) (18 vs. 19h; p<0.001). In relation to patients with MV duration >48h, those in phase II had a comparatively higher successful extubation rate (p=0.033), shorter ICU LOS (p=0.038) and a significant decrease in in-hospital mortality (p=0.039). CONCLUSIONS: The intensivist-directed care model showed improved rates of successful extubation and shorter MV durations after cardiac surgery.
Assuntos
Extubação/estatística & dados numéricos , Procedimentos Cirúrgicos Cardíacos , Cuidados Críticos/métodos , Mortalidade Hospitalar , Cuidados Pós-Operatórios/métodos , Respiração Artificial/estatística & dados numéricos , Adulto , Estudos de Casos e Controles , China , Unidades de Cuidados Coronarianos , Feminino , Humanos , Intubação/estatística & dados numéricos , Tempo de Internação , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , Cirurgiões , Fatores de TempoRESUMO
OBJECTIVES: Degenerative disc disease (DDD) and osteoarthritis (OA) are relatively frequent causes of disability amongst the elderly; they constitute serious socioeconomic costs and significantly impair quality of life. Previous studies to date have found that aggrecan variable number of tandem repeats (VNTR) contributes both to DDD and OA. However, current data are not consistent across studies. The purpose of this study was to evaluate systematically the relationship between aggrecan VNTR, and DDD and/or OA. METHODS: This study used a highly sensitive search strategy to identify all published studies related to the relationship between aggrecan VNTR and both DDD and OA in multiple databases from January 1996 to December 2016. All identified studies were systematically evaluated using specific inclusion and exclusion criteria. Cochrane methodology was also applied to the results of this study. RESULTS: The final selection of seven studies was comprehensively evaluated and includes results for 2928 alleles. The most frequent allele among all the studies was allele 27. After comparing the distributions of each allele with others, statistically significant differences have been found in the distribution of the alleles by the two groups, with an over-representation of allele (A)21 (disease: 3.22%, control: 0.44%). Thus, carrying A21 increased the risk of DDD. Such an association was not found to be statistically significant when considering the risk of OA. CONCLUSIONS: The findings suggest that VNTR A21 seems to be associated with higher risk to DDD, however, such an association may not be statistically significant regarding the risk of OA.Cite this article: L. Cong, G. Tu, D. Liang. A systematic review of the relationship between the distributions of aggrecan gene VNTR polymorphism and degenerative disc disease/osteoarthritis. Bone Joint Res 2018;7:308-317. DOI: 10.1302/2046-3758.74.BJR-2017-0207.R1.
RESUMO
Ortleppascaris sinensis (Nematoda: Ascaridida) is a dominant intestinal nematode of the captive Chinese alligator. However, the epidemiology, molecular ecology and population genetics of this parasite remain largely unexplored. In this study, the complete mitochondrial (mt) genome sequence of O. sinensis was first determined using a polymerase chain reaction (PCR)-based primer-walking strategy, and this is also the first sequencing of the complete mitochondrial genome of a member of the genus Ortleppascaris. The circular mitochondrial genome (13,828 bp) of O. sinensis contained 12 protein-coding, 22 transfer RNA and 2 ribosomal RNA genes, but lacked the ATP synthetase subunit 8 gene. Finally, phylogenetic analysis of mtDNAs indicated that the genus Ortleppascaris should be attributed to the family Heterocheilidae. It is necessary to sequence more mtNDAs of Ortleppascaris nematodes in the future to test and confirm our conclusion. The complete mitochondrial genome sequence of O. sinensis reported here should contribute to molecular diagnosis, epidemiological investigations and ecological studies of O. sinensis and other related Ascaridida nematodes.
Assuntos
Ascaridoidea/genética , DNA Mitocondrial/genética , Genoma Helmíntico , Genoma Mitocondrial , Jacarés e Crocodilos/parasitologia , Animais , Ascaridoidea/classificação , Ascaridoidea/isolamento & purificação , Sequenciamento de Nucleotídeos em Larga Escala , Intestinos/parasitologia , Filogenia , Reação em Cadeia da Polimerase , RNA Ribossômico , RNA de Transferência , Análise de Sequência de DNARESUMO
Objective: To discuss the strategy of therapeutic management of T3 supraglottic carcinoma. Methods: A retrospective analysis of 459 patients with T3 supraglottic carcinoma treated in our hospital was performed. We evaluated the results of different managements, including surgery alone, preoperative radiotherapy, postoperative radiotherapy and radiotherapy alone. The extent of the lesion was also put into analysis. Statistical analysis of the overall survival (OS), cause-specific survival (CSS), local control (LC), regional control(RC), function-conservation (FC) were performed with the statistical package from SPSS. Results: In all patients, the rates of 5-year OS, CSS, LC, RC and FC were 64.2%, 71.2%, 87.8%, 78.8% and 64.5% respectively. The OS, LC and FC of the patients treated by surgery alone, preoperative radiotherapy and postoperative radiotherapy had no significant difference, and were remarkably better than that of patients treated by radiotherapy alone (P<0.001). In 412 patients treated by surgery, 300 patients received function-conservation laryngectomy. 209 patients (50.7%, 209/412) survived and maintained well-function of larynx for 5 years, which was significantly better than those in the radiotherapy alone group (27.7%, 13/47). The patients with the lesion invading the pre-epiglottic space but limited in supraglottic area had better OS (70.2%), LC (93.5%) and FC (85.1%). The rate of 5-year neck lymphatic metastasis was 56.2%(258/459), and the 5-year OS of patients with N0, N1, N2 and N3 stage were 76.0%, 66.2%, 50.5% and 13.0% respectively. Conclusions: Surgical treatment was the best therapeutic approach for T3 supraglottic laryngeal carcinoma. Most patients with T3 lesions are suitable for function-conservation laryngectomy. Surgical procedure was determined by tumor invaded location and extension. The combined therapy of surgery and radiotherapy had no significant advantage.
Assuntos
Carcinoma/radioterapia , Carcinoma/cirurgia , Neoplasias Laríngeas/radioterapia , Neoplasias Laríngeas/cirurgia , Carcinoma/mortalidade , Carcinoma/patologia , Humanos , Neoplasias Laríngeas/mortalidade , Neoplasias Laríngeas/patologia , Laringectomia/métodos , Laringe , Metástase Linfática , Pescoço , Estadiamento de Neoplasias , Tratamentos com Preservação do Órgão , Radioterapia Adjuvante , Estudos Retrospectivos , Análise de SobrevidaRESUMO
OBJECTIVE: This study aimed to evaluate the outcomes of high-flow nasal cannula (HFNC) oxygen therapy compared with noninvasive ventilation (NIV) for the treatment of acute hypoxemic respiratory failure in renal transplant recipients. METHODS: Data were retrospectively collected from a tertiary intensive care unit (ICU) from July 1, 2011, to September 31, 2015. All renal recipients who had acute respiratory failure at that period of time were classified into the HFNC or NIV group depending on the initial form of respiratory support. RESULTS: A total of 38 patients were enrolled in this study. Twenty patients received HFNC and the other 18 received NIV as the initial respiratory support. The ICU mortality in the HFNC group was 5% (1 patient), compared with 22.2% (4 patients) in the NIV group (P = .083). The median length of the ICU stay was 12 days in the HFNC group, compared with 14 days in the NIV group (P = .297). The number of ventilator-free days at day 28 was significantly higher in the HFNC group than in the NIV group (26 ± 3 vs 21 ± 3; P < .001). The incidences of both pneumothorax (0% vs 22.2%; P = .042) and skin breakdown (0% vs 22.2%; P = .042) were significantly lower in the HFNC group. CONCLUSIONS: In renal transplant recipients with acute hypoxemic respiratory failure secondary to severe pneumonia, HFNC achieved outcomes similar to NIV. In addition, HFNC was associated with an increased number of ventilator-free days at day 28 and fewer complications.
Assuntos
Cânula , Hipóxia/terapia , Transplante de Rim/efeitos adversos , Ventilação não Invasiva/métodos , Oxigenoterapia/métodos , Complicações Pós-Operatórias , Insuficiência Respiratória/terapia , Doença Aguda , Feminino , Humanos , Hipóxia/etiologia , Unidades de Terapia Intensiva , Masculino , Pessoa de Meia-Idade , Oxigenoterapia/instrumentação , Insuficiência Respiratória/etiologia , Estudos Retrospectivos , Resultado do TratamentoRESUMO
Endothelin-1 (ET-1) is the most potent endogenous vasoconstrictor and is involved in several vascular disorders such as hypertension. Its strong interaction with other vasoactive hormone systems suggests that the ET-1 gene (EDN1) is a potential candidate molecule that influences the risk of developing hypertension. Recently, two single nucleotide polymorphisms in EDN1 have been reported to be associated with hypertension: Lys198Asn and 3A/4A (-134delA) located in the 5'-untranslated region. To determine the association of these two polymorphisms with hypertension, we genotyped patients and controls (N = 537) and compared the allele and genotype frequencies between groups. There was no significant difference in the genotype frequencies of these two polymorphisms between healthy controls and hypertension patients. Although previous reports have revealed a significant interaction between the END1 Lys198Asn (G/T) polymorphism and body mass index in association with hypertension, no such relationship was observed in the present study. Further, we compared blood pressure among hypertensive subjects and observed that neither systolic nor diastolic blood pressure was significantly associated with variations in the genotypes of the two single nucleotide polymorphisms. In summary, these two END1 polymorphisms do not appear to affect the development of hypertension in the Chinese population.
Assuntos
Endotelina-1/genética , Hipertensão/genética , Polimorfismo de Nucleotídeo Único , Idoso , Estudos de Casos e Controles , China , Feminino , Frequência do Gene , Humanos , Masculino , Pessoa de Meia-Idade , Mutação de Sentido IncorretoRESUMO
OBJECTIVE: To evaluate the underlying function of microRNAs (miRNAs) in osteoarthritis (OA). DESIGN: A bioinformatic analysis of miRNAs-OA studies was completed in multiple databases. All identified articles were assessed using specific inclusion and exclusion criteria (Eligible case-control studies for the present study included those which investigated miRNAs differential expression in cartilage tissues and cells of OA and controls. Abstracts, case reports, conference presentations, editorials, and expert opinions were excluded.). We performed bioinformatic analysis and assessed which miRNAs are commonly elevated or decreased in cartilage of OA, and assessed putative targets of these miRNAs using TargetScan, Database for Annotation, Visualization and Integrated Discovery (DAVID), FunRich and String. RESULTS: Fifty seven studies were included in this study. Our current review has identified 46 differentially expressed miRNAs involved in autophagy, inflammation, chondrocyte apoptosis, chondrocyte differentiation & homeostasis, chondrocyte metabolism and degradation of the extracellular matrix (ECM). Additionally, our literature search identified a wide range of miRNAs that have been shown to be differentially expressed in OA. The function of up-regulated miRNAs primarily target nucleus, whereas the function of down-regulated miRNAs primarily target transcription. CONCLUSIONS: Comprehensive analysis of all miRNAs studies reveals cooperation in miRNA signatures and suggests that there may be two biologically synergic classes of miRNAs that are associated with OA. This finding suggests that miRNAs may be useful as diagnostic biomarkers and/or may provide new therapeutic targets in OA. Furthermore, a better understanding of the targets of these miRNAs will accelerate biomedical discoveries and improve clinical care based on new knowledge of OA-related disease mechanisms.
Assuntos
Cartilagem Articular/fisiologia , MicroRNAs/fisiologia , Osteoartrite/etiologia , Apoptose/fisiologia , Autofagia/fisiologia , Diferenciação Celular/fisiologia , Condrócitos/fisiologia , Biologia Computacional , Homeostase/fisiologia , Humanos , MicroRNAs/metabolismo , Osteoartrite/fisiopatologia , Osteocondrite/fisiopatologiaRESUMO
Multiple drug resistance is a challenging issue in the clinic. There is growing evidence that the G-protein-coupled estrogen receptor (GPER) is a novel mediator in the development of multidrug resistance in both estrogen receptor (ER)-positive and -negative breast cancers, and that cancer-associated fibroblasts (CAFs) in the tumor microenvironment may be a new agent that promotes drug resistance in tumor cells. However, the role of cytoplasmic GPER of CAFs on tumor therapy remains unclear. Here we first show that the breast tumor cell-activated PI3K/AKT (phosphoinositide 3-kinase/AKT) signaling pathway induces the cytoplasmic GPER translocation of CAFs in a CRM1-dependent pattern, and leads to the activation of a novel estrogen/GPER/cAMP/PKA/CREB signaling axis that triggers the aerobic glycolysis switch in CAFs. The glycolytic CAFs feed the extra pyruvate and lactate to tumor cells for augmentation of mitochondrial activity, and this energy metabolically coupled in a 'host-parasite relationship' between catabolic CAFs and anabolic cancer cells confers the tumor cells with multiple drug resistance to several conventional clinical treatments including endocrine therapy (tamoxifen), Her-2-targeted therapy (herceptin) and chemotherapy (epirubicin). Moreover, the clinical data from 18F-fluorodeoxyglucose positron emission tomography/computed tomography further present a strong association between the GPER/cAMP/PKA/CREB pathway of stromal fibroblasts with tumor metabolic activity and clinical treatment, suggesting that targeting cytoplasmic GPER in CAFs may rescue the drug sensitivity in patients with breast cancer. Thus, our data define novel insights into the stromal GPER-mediated multiple drug resistance from the point of reprogramming of tumor energy metabolism and provide the rationale for CAFs as a promising target for clinical therapy.
Assuntos
Neoplasias da Mama/metabolismo , Proteína de Ligação ao Elemento de Resposta ao AMP Cíclico/metabolismo , Proteínas Quinases Dependentes de AMP Cíclico/metabolismo , AMP Cíclico/metabolismo , Receptores de Estrogênio/metabolismo , Receptores Acoplados a Proteínas G/metabolismo , Transporte Ativo do Núcleo Celular , Neoplasias da Mama/patologia , Linhagem Celular Tumoral , Citoplasma/metabolismo , Resistencia a Medicamentos Antineoplásicos , Feminino , Fibroblastos/metabolismo , Fibroblastos/patologia , Glicólise , Humanos , Carioferinas/metabolismo , Células MCF-7 , Proteínas Proto-Oncogênicas c-akt/metabolismo , Receptores Citoplasmáticos e Nucleares/metabolismo , Células Estromais/metabolismo , Células Estromais/patologia , Proteína Exportina 1RESUMO
Some insect chitinases are required to degrade chitin and ensure successful metamorphosis. Although chitinase genes have been well characterized in several model insects, no reports exist for the rice striped stem borer, Chilo suppressalis, a highly destructive pest that causes huge yield losses in rice production. Here, we conducted a genome-level analysis of chitinase genes in C. suppressalis. After amplification of full-length transcripts with rapid amplification of cDNA ends, we identified 12 chitinase genes in C. suppressalis. All these genes had the conserved domains and motifs of glycoside hydrolase family 18 and grouped phylogenetically into five subgroups. C. suppressalis chitinase 1 (CsCht1) was highly expressed in late pupae, whereas CsCht3 was abundant in early pupae. Both CsCht2 and CsCht4 were highly expressed in larvae. CsCht2 was abundant specifically in the third-instar larvae and CsCht4 showed periodic high expression in 2- to 5-day-old larvae in each instar. Tissue specific expression analysis indicated that CsCht1 and CsCht3 were highly expressed in epidermis whereas CsCht2 and CsCht4 were specifically abundant in the midgut. Knockdown of CsCht1 resulted in adults with curled wings, indicating that CsCht1 might have an important role in wing expansion. Silencing of CsCht2 or CsCht4 arrested moulting, suggesting essential roles in larval development. When the expression of CsCht3 was interfered, defects in pupation occurred. Overall, we provide here the first catalogue of chitinase genes in the rice striped stem borer and have elucidated the functions of four chitinases in metamorphosis.
Assuntos
Quitinases/genética , Genoma de Inseto , Metamorfose Biológica , Muda , Mariposas/genética , Animais , Quitinases/metabolismo , Larva/enzimologia , Larva/genética , Larva/crescimento & desenvolvimento , Mariposas/enzimologia , Mariposas/crescimento & desenvolvimento , Filogenia , Pupa/enzimologia , Pupa/genética , Pupa/crescimento & desenvolvimento , Análise de Sequência de DNARESUMO
A new nematode species, Ortleppascaris sinensis sp. nov. (Ascaridoidea), is described from specimens found in the stomach and intestine of the Chinese alligator Alligator sinensis Fauvel, 1879 (Crocodilian: Alligatoridae) in the National Nature Reserve of Chinese Alligator (Chinese Crocodile Lake) in Anhui Province, China. This is the first description of O. sinensis sp. nov. in both China and this crocodile host, increasing its distribution in South Asia as well as expanding the number of helminths known to infect this crocodile. The detailed description of O. sinensis sp. nov., based on light and scanning electron microscopic examination, provides new taxonomic data for this species, and we also report sequences of the internal transcribed spacers (ITS), small subunit DNA segments (18S) and the cytochrome oxidase I (COI) gene.
Assuntos
Jacarés e Crocodilos/parasitologia , Infecções por Ascaridida/veterinária , Ascaridoidea/anatomia & histologia , Ascaridoidea/genética , Animais , Infecções por Ascaridida/parasitologia , Ascaridoidea/classificação , Ásia , China , DNA de Helmintos/química , DNA de Helmintos/genética , DNA Espaçador Ribossômico/química , DNA Espaçador Ribossômico/genética , Feminino , Intestinos/parasitologia , Masculino , Microscopia , Análise de Sequência de DNA , Estômago/parasitologiaRESUMO
BACKGROUND: Non-pharmacological interventions are effective neonatal pain reduction strategies. We aimed to study the effects of non-nutritive sucking (NNS) and swaddling on infants' behavioural and physiological parameters during shallow or deep heel stick procedures. METHOD: In this prospective, multi-centred, randomized controlled clinical trial, we enrolled 671 newborns. The infants undergoing shallow or deep heel stick procedures were randomized into four groups: oral sucrose (routine care, group S), oral sucrose combined with NNS (group NS), oral sucrose combined with swaddling (group SS) and oral sucrose combined with NNS and swaddling (group NSS). The behavioural responses were evaluated by the Revised Neonatal Facial Coding System and the physiological signals were monitored by electrocardiogram monitors. RESULTS: A significant synergistic analgesic effect was observed between the NS and SS groups in both the shallow (F = 5.952, p = 0.015) and deep heel stick (F = 7.452, p = 0.007) procedure. NSS group exhibited the lowest pain score. For the deep heel stick procedure, the NS group had a significantly lower increase in heart rate (HR)% and decrease in SPO2 % than the S group (F = 17.540, p = 0.000, F = 10.472, p = 0.001), while this difference was not observed in the shallow heel stick procedure. No difference was found between the S and SS groups, in terms of different physiological parameters. CONCLUSION: Non-nutritive sucking and swaddling had synergistic effects on pain relief when used with oral sucrose. For the deep heel stick procedure, oral sucrose combined with NNS and swaddling provided the best pain relief effect. For the shallow heel stick procedure, addition of NNS and swaddling did not improve the effects.
Assuntos
Coleta de Amostras Sanguíneas/efeitos adversos , Manejo da Dor/métodos , Dor Processual/terapia , Feminino , Frequência Cardíaca , Humanos , Comportamento do Lactente , Cuidado do Lactente , Recém-Nascido , Masculino , Medição da Dor , Dor Processual/diagnóstico , Dor Processual/etiologia , Estudos Prospectivos , Comportamento de Sucção , Sacarose/uso terapêutico , Edulcorantes/uso terapêuticoRESUMO
Based on the observation of a grey phenotype in the F(1) generation from a cross between two white plumage duck varieties, the white Kaiya and the white Liancheng, we hypothesized a possible interaction between two autosomal loci that determine grey plumage. Using the parental and F(1) individuals, seven testing combinations including five different F(1) intercrosses (F(2)) and two different backcrosses (BC(1) and BC(2)) were designed to test our hypothesis. It was demonstrated by chi-squared analysis that six test matings produced offspring in the expected ratios between the grey and white, with P-values ranging from 0.50 to 0.99. Another mating, where all white offspring were expected, produced 33 white individuals. These results verified that the interaction between two loci produced the grey phenotype. The C locus, which carries the recessive allele (c), was previously thought to be the only gene responsible for white plumage in the duck. This is the first report that an allele (t), carried by the white Liancheng at a different autosomal locus, also determines white plumage in ducks. Furthermore, the dominant alleles at both loci can interact with each other to produce the grey phenotype, and a new dark phenotype, observed in some F(2) individuals, can be attributed to the dosage effect of the T allele.
Assuntos
Patos/anatomia & histologia , Patos/genética , Pigmentação/genética , Animais , Cruzamentos Genéticos , Epistasia Genética , Genes Dominantes , Genes RecessivosRESUMO
The G-protein-coupled-receptor 30 (GPR30) is a new membrane estrogen receptor. The aim of the present study was to determine the correlations among GPR30, ERalpha, PR, C-erbB-2, p53, TNM stage, and pathologic grade in breast carcinomas. Two hundred forty-one biopsy specimens were evaluated with immunohistochemical assays, and then correlations were analyzed. Low negative correlations of GPR30 with ERalpha (r = -0.144, P<0.05) and PR (r = -0.214, P<0.01) were observed. Associations of GPR30 with C-erbB-2, p53, TNM stage, and pathologic grade were not confirmed. These findings indicated that GPR30 might be an independent prognostic factor in breast carcinomas.
Assuntos
Neoplasias da Mama/química , Neoplasias da Mama/patologia , Receptores Acoplados a Proteínas G/análise , Receptor alfa de Estrogênio/análise , Feminino , Humanos , Imuno-Histoquímica , Receptor ErbB-2/análise , Receptores de Estrogênio , Receptores de Progesterona/análiseRESUMO
Enterohemorrhagic Escherichia coli (EHEC) O157:H7 intimately attaches to intestinal epithelial monolayers and produces attaching and effacing (A/E) lesions. In addition, EHEC infection causes disruptions of intercellular tight junctions, leading to clinical sequelae that include acute diarrhea, hemorrhagic colitis, and the hemolytic-uremic syndrome. Current therapy remains supportive since antibiotic therapy increases the risk of systemic complications. This study focused on the potential therapeutic effect of an alternative form of therapy, probiotic Lactobacillus rhamnosus strain GG, to attenuate EHEC-induced changes in paracellular permeability in polarized MDCK-I and T84 epithelial cell monolayers. Changes in epithelial cell morphology, electrical resistance, dextran permeability, and distribution and expression of claudin-1 and ZO-1 were assessed using phase-contrast, immunofluorescence, and transmission electron microscopy and macromolecular flux. This study demonstrated that pretreatment of polarized MDCK-I and T84 cells with the probiotic L. rhamnosus GG reduced morphological changes and diminished the number of A/E lesions induced in response to EHEC O157:H7 infection. With probiotic pretreatment there was corresponding attenuation of the EHEC-induced drop in electrical resistance and the increase in barrier permeability assays. In addition, L. rhamnosus GG protected epithelial monolayers against EHEC-induced redistribution of the claudin-1 and ZO-1 tight junction proteins. In contrast to the effects seen with the live probiotic, heat-inactivated L. rhamnosus GG had no effect on EHEC binding and A/E lesion formation or on disruption of the barrier function. Collectively, these findings provide in vitro evidence that treatment with the probiotic L. rhamnosus strain GG could prove to be an effective management treatment for preventing injury of the epithelial cell barrier induced by A/E bacterial enteropathogens.
Assuntos
Células Epiteliais/microbiologia , Células Epiteliais/fisiologia , Escherichia coli O157/fisiologia , Lacticaseibacillus rhamnosus/classificação , Lacticaseibacillus rhamnosus/fisiologia , Animais , Aderência Bacteriana/fisiologia , Linhagem Celular , Claudina-1 , Cães , Células Epiteliais/patologia , Regulação da Expressão Gênica , Proteínas de Membrana/genética , Proteínas de Membrana/metabolismo , Permeabilidade , Fosfoproteínas/genética , Fosfoproteínas/metabolismo , Transporte Proteico , Proteína da Zônula de Oclusão-1RESUMO
Two new arylbenzofurans, hedysarimbenzofuran A (1) and hedysarimbenzofuran B (2), were isolated from the roots of Hedysarum multijugum. The structures were determined by spectroscopic methods.
Assuntos
Benzofuranos/isolamento & purificação , Fabaceae/química , Benzofuranos/química , Estrutura Molecular , Ressonância Magnética Nuclear Biomolecular , Raízes de Plantas/química , Espectrometria de Massas por Ionização por Electrospray , Espectrofotometria UltravioletaRESUMO
Previous experiments have demonstrated the stable expression of factor IX (FIX) protein in mice and canine models of hemophilia B following portal vein gene transfer with a recombinant adeno-associated virus (rAAV) vector encoding FIX. Here, we present the results of studies that further optimized the rAAV vector transgene cassette used to express FIX and explored the use of the less-invasive intravenous (i.v.) route of vector administration for the treatment of hemophilia B. First, a liver-specific promoter was evaluated in conjunction with cis-acting regulatory elements in mice. Constructs that included both the beta-globin intron and the woodchuck hepatitis virus post-transcriptional regulatory element resulted in the highest level of FIX expression in vivo. Using this optimized vector, we demonstrate that i.v. injection was feasible for hepatic gene transfer in mice, achieving 70-80% of portal vein expression levels of FIX. In further studies using the Chapel Hill strain of hemophilia B dogs, we demonstrate for the first time FIX expression and partial correction of the bleeding disorder following i.v. administration of an AAV vector.
Assuntos
Dependovirus/genética , Fator IX/genética , Terapia Genética/métodos , Vetores Genéticos/administração & dosagem , Hemofilia B/terapia , Animais , Cães , Fator IX/análise , Expressão Gênica , Vetores Genéticos/genética , Veias Hepáticas , Humanos , Infusões Intravenosas , Injeções Intravenosas , Fígado/metabolismo , Camundongos , Cauda/irrigação sanguínea , Transdução Genética/métodosRESUMO
This article represents the proceedings of a symposium at the 2001 annual meeting of the Research Society on Alcoholism in Montreal, Canada. Drs. Yedy Israel and Fulton Crews were organizers and co-chairpersons. The presentations were (1) Introduction to the symposium, by Yedy Israel; (2) Gene delivery to the brain, by Fulton T. Crews; (3) Gene therapy in alcoholic liver injury, by Ronald Thurman; and (4) Antisense oligonucleotides and antisense-gene delivery, by Yedy Israel.