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Diabetes is a chronic condition that has reached epidemic proportions in the United States, affecting nearly 34 million adults, and disproportionately affecting vulnerable populations, such as ethnic minorities, the elderly and individuals with low socioeconomic status. This study addresses the impact of the Health Extension for Diabetes (HED) program, a community-based diabetes self-management support program, on adult diabetes self-care behaviors. The Summary of Diabetes Self-Care Activities (SDSCA) was utilized to evaluate improvement in diabetes self-care behaviors. Descriptive statistics, univariate and multivariable regression models were conducted. Significant increases were observed among program participants (N = 149) in all five subscales of the SDSCA (general diet, specific diet, blood glucose testing, exercise and foot care; P-values < 0.001). A priority of this diabetes education program was helping underserved populations; over half (62%) of participants self-identified as Black/African Americans. After program participation, scores on all SDSCA subscales increased significantly among Black/African Americans (n = 93) by approximately 1 day per week. White/other races (n = 56) showed similar increases in four of the SDSCA subscales post-HED program participation. This study shows that increasing participation in community-based, diabetes self-management support programs, such as HED, can increase engagement in diabetes self-care behaviors among underserved groups.
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Diabetes Mellitus Tipo 2 , Diabetes Mellitus , Autogestão , Humanos , Adulto , Estados Unidos , Idoso , Autocuidado , Diabetes Mellitus/terapia , Comportamentos Relacionados com a Saúde , Dieta , Diabetes Mellitus Tipo 2/terapiaRESUMO
Mapping of the protein structural flexibility with sub-2-nm spatial resolution in liquid is achieved by combining bimodal excitation and frequency modulation force microscopy. The excitation of two cantilever eigenmodes in dynamic force microscopy enables the separation between topography and flexibility mapping. We have measured variations of the elastic modulus in a single antibody pentamer from 8 to 18 MPa when the probe is moved from the end of the protein arm to the central protrusion. Bimodal dynamic force microscopy enables us to perform the measurements under very small repulsive loads (30-40 pN).
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Módulo de Elasticidade , Microscopia de Força Atômica/instrumentação , Proteínas/química , ElasticidadeRESUMO
INTRODUCTION: Microgravity has been shown to impose various effects on breast cancer cells. We exposed human breast cancer cells to simulated microgravity and studied morphology and alterations in gene expression. MATERIALS AND METHODS: Human breast cancer cells were exposed to simulated microgravity in a random positioning machine (RPM) for 24 h. Morphology was observed under light microscopy, and gene alteration was studied by qPCR. RESULTS: After 24 h, formation of three-dimensional structures (spheroids) occurred. BRCA1 expression was significantly increased (1.9×, p < 0.05) in the adherent cells under simulated microgravity compared to the control. Expression of KRAS was significantly decreased (0.6×, p < 0.05) in the adherent cells compared to the control. VCAM1 was significantly upregulated (6.6×, 2.0×, p < 0.05 each) in the adherent cells under simulated microgravity and in the spheroids. VIM expression was significantly downregulated (0.45×, 0.44×, p < 0.05 each) in the adherent cells under simulated microgravity and in the spheroids. There was no significant alteration in the expression of MAPK1, MMP13, PTEN, and TP53. CONCLUSIONS: Simulated microgravity induces spheroid formation in human breast cancer cells within 24 h and alters gene expression toward modified adhesion properties, enhanced cell repair, and phenotype preservation. Further insights into the underlying mechanisms could open up the way toward new therapies.
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Neoplasias da Mama/genética , Neoplasias da Mama/fisiopatologia , Expressão Gênica/genética , Simulação de Ausência de Peso/efeitos adversos , Ausência de Peso/efeitos adversos , Linhagem Celular Tumoral , Feminino , Expressão Gênica/fisiologia , Humanos , Fenótipo , Transdução de Sinais/genética , Transdução de Sinais/fisiologia , Regulação para Cima/genética , Regulação para Cima/fisiologiaRESUMO
Three-dimensional plasma crystals are often described as Yukawa systems for which a phase transition between the crystal structures fcc and bcc has been predicted. However, experimental investigations of this transition are missing. We use a fast scanning video camera to record the crystallization process of 70 000 microparticles and investigate the existence of the fcc-bcc phase transition at neutral gas pressures of 30, 40, and 50 Pa. To analyze the crystal, robust phase diagrams with the help of a machine learning algorithm are calculated. This work shows that the phase transition can be investigated experimentally and makes a comparison with numerical results of Yukawa systems. The phase transition is analyzed in dependence on the screening parameter and structural order. We suggest that the transition is an effect of gravitational compression of the plasma crystal. Experimental investigations of the fcc-bcc phase transition will provide an opportunity to estimate the coupling strength Γ by comparison with numerical results of Yukawa systems.
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In this work, we report on the small scale polycondensation and consecutive analysis of novel polyesters based on the potentially renewable 1,3-cyclopentanediol (CPdiol). To avoid evaporation of monomers during thin-film polymerization reactions, trimer pre-polyesters have been synthesized from the corresponding acid-chlorides with diol monomers. Polymerization of these trimers was explored by thermogravimetric analysis to identify potential side reactions, and to assess the ideal polymerization temperature. In general we observe that trans-1,3-cyclopentanediol exhibits good thermal stability up to 200 °C, whereas thermal dehydration of the alcohol end-groups occurs upon further heating. In contrast, for cis-1,3-cyclopentanediol, the ester bonds of the cyclopentane end-groups become labile, thereby generating carboxylic acid end-groups, and 3-cyclopentenol already at 180 °C. The thermal dehydration reactions yield double bond end-groups, which in turn facilitate cross-linking through cross-coupling and Diels-Alder reactions, leading to an increase in molecular weight. Despite the limited thermal stability of CPdiol, here we demonstrate that polymerization of CPdiol can successfully be achieved in thin-film polycondensation conditions at 180 °C, yielding molecular weights well above 10 kg mol-1.
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This corrects the article DOI: 10.1038/leu.2017.9.
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The feasibility of miniaturised pressurised liquid extraction (PLE) with in-cell purification and subsequent gas chromatography with micro-electron capture detection (GC-micro-ECD) for the determination of prioritary and toxic polychlorinated biphenyls (PCBs) in a variety of foodstuffs (fat contents in the range 22-49%, w/w, on a freeze-dried basis) has been investigated. After optimisation of the several experimental parameters affecting the efficiency of the selective PLE process, the developed method provided quantitative recoveries of the endogenous PCBs studied and complete fat elimination in a single step using n-hexane as extraction solvent. A total solvent volume of 3.5 mL was used for the two consecutive 7 min static PLEs of 100-mg samples. Detection limits using GC-micro-ECD were below 0.2 ng/g freeze dried sample for all 22 PCBs investigated in real-life foodstuffs, and the repeatability of the complete PLE plus GC-micro-ECD method as calculated for the analysis of the endogenous PCBs in general was better than 14%. Comparison of the miniaturised PLE method developed with either conventional Soxhlet extraction or matrix solid phase dispersion with subsequent (off-line) clean-up for the analysis of non-spiked samples showed that the efficiency of PLE was similar to or better (recoveries in the range 83-133%, as calculated for the endogenous analytes) than for the other two extraction methods assayed.
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Métodos Analíticos de Preparação de Amostras/métodos , Contaminação de Alimentos/análise , Bifenilos Policlorados/análise , Cromatografia Gasosa/métodos , Gorduras na Dieta/análise , Miniaturização/métodos , PressãoRESUMO
An approach for automated nanotomography, a layer-by-layer imaging technique based on scanning probe microscopy (SPM), is presented. Stepwise etching and imaging is done in situ in a liquid cell of an SPM. The flow of etching and rinsing solutions after each etching step is controlled with solenoid valves which allow for an automated measuring protocol. The thermal drift and the drift of the piezo scanner is corrected by applying offsets calculated from the cross correlation coefficients between successive images. As an example, we have imaged human bone with approximately 10 nm resolution using tapping mode SPM and successive etching with hydrochloric acid.
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Aumento da Imagem/instrumentação , Microscopia de Varredura por Sonda/instrumentação , Nanoestruturas/ultraestrutura , Nanotecnologia/instrumentação , Robótica/instrumentação , Manejo de Espécimes/instrumentação , Tomografia Óptica/instrumentação , Aumento da Imagem/métodos , Microscopia de Varredura por Sonda/métodos , Nanotecnologia/métodos , Reprodutibilidade dos Testes , Robótica/métodos , Sensibilidade e Especificidade , Manejo de Espécimes/métodos , Tomografia Óptica/métodosRESUMO
Machine learning is one of the most popular fields in computer science and has a vast number of applications. In this work we will propose a method that will use a neural network to locally identify crystal structures in a mixed phase Yukawa system consisting of fcc, hcp, and bcc clusters and disordered particles similar to plasma crystals. We compare our approach to already used methods and show that the quality of identification increases significantly. The technique works very well for highly disturbed lattices and shows a flexible and robust way to classify crystalline structures that can be used by only providing particle positions. This leads to insights into highly disturbed crystalline structures.
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A sample holder with a large open area offers several benefits when used in the process of plasma immersion ion implantation and deposition in which the plasma is generated by a high voltage applied to the sample holder: The ignition voltage of the plasma is lower, and the deposition rate can be several times higher than in the case of a normal plate-like holder. There is a more pronounced edge effect regarding the film thickness. Other film properties are also affected; for diamond-like carbon films, the film structure exhibits more disorder. The hardness of the samples is similar, with the surfaces of the samples being very smooth.
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It is unknown, why only a minority of chronic myeloid leukemia (CML) patients sustains treatment free remission (TFR) after discontinuation of tyrosine kinase inhibitor (TKI) therapy in deep molecular remission (MR). Here we studied, whether expression of the T-cell inhibitory receptor (CTLA-4)-ligand CD86 (B7.2) on plasmacytoid dendritic cells (pDC) affects relapse risk after TKI cessation. CML patients in MR displayed significantly higher CD86+pDC frequencies than normal donors (P<0.0024), whereas TFR patients had consistently low CD86+pDC (n=12). This suggested that low CD86+pDC might be predictive of TFR. Indeed, in a prospective analysis of 122 patients discontinuing their TKI within the EURO-SKI trial, the one-year relapse-free survival (RFS) was 30.1% (95% CI 15.6-47.9) for patients with >95 CD86+pDC per 105 lymphocytes, but 70.0% (95% CI 59.3-78.3) for patients with <95 CD86+pDC (hazard ratio (HR) 3.4, 95%-CI: 1.9-6.0; P<0.0001). Moreover, only patients with <95 CD86+pDC derived a significant benefit from longer (>8 years) TKI exposure before discontinuation (HR 0.3, 95% CI 0.1-0.8; P=0.0263). High CD86+pDC counts significantly correlated with leukemia-specific CD8+ T-cell exhaustion (Spearman correlation: 0.74, 95%-CI: 0.21-0.92; P=0.0098). Our data demonstrate that CML patients with high CD86+pDC counts have a higher risk of relapse after TKI discontinuation.
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Antígeno B7-2/metabolismo , Antígeno CTLA-4/metabolismo , Células Dendríticas/metabolismo , Leucemia Mielogênica Crônica BCR-ABL Positiva/metabolismo , Adulto , Idoso , Antígeno B7-2/genética , Biomarcadores , Contagem de Células , Células Dendríticas/imunologia , Feminino , Expressão Gênica , Humanos , Imunofenotipagem , Estimativa de Kaplan-Meier , Leucemia Mielogênica Crônica BCR-ABL Positiva/diagnóstico , Leucemia Mielogênica Crônica BCR-ABL Positiva/tratamento farmacológico , Leucemia Mielogênica Crônica BCR-ABL Positiva/mortalidade , Masculino , Pessoa de Meia-Idade , Prognóstico , Inibidores de Proteínas Quinases/uso terapêutico , Recidiva , Indução de Remissão , Subpopulações de Linfócitos T/imunologia , Subpopulações de Linfócitos T/metabolismo , Resultado do Tratamento , Adulto JovemRESUMO
The objective of this study was to elucidate the role of uridine for spermatozoa, since this pyrimidine nucleoside was found in millimolar concentration in human seminal plasma. Here, the degradative activity of uridine-phosphorylase [EC 2.4.2.3] and the salvage activity of uridine kinase [EC 2.7.1.48] were detected in human spermatozoa. HPLC analysis depicted the uptake of exogeneous 14C-labelled adenine, but not of uridine and of hypoxanthine, into nucleotide pools of boar spermatozoa. On addition of uridine, the computer-assisted semen analysis (CASA) of human cells revealed a reduction of the percentage of motile spermatozoa in contrast to an elevation of some velocity parameters. It is concluded that exogeneous uridine could function as suppressor for early capacitation and as a substrate for phosphorolysis, if ribose is needed, rather than to satisfy a demand for intracellular pyrimidine nucleotides.
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Espermatozoides/metabolismo , Espermatozoides/ultraestrutura , Uridina/química , Uridina/metabolismo , Adenina/química , Animais , Cromatografia Líquida de Alta Pressão , Humanos , Hipoxantina/metabolismo , Processamento de Imagem Assistida por Computador , Masculino , Nucleotídeos/química , Fosforilação , Ribose/química , Sêmen/metabolismo , Motilidade dos Espermatozoides , Espermatozoides/patologia , Uridina Fosforilase/fisiologiaRESUMO
In this work, existing methods for plasma crystal analysis are investigated using artificial and simulated calibration data, which reproduce a multiphase system consisting of fcc, hcp, and bcc crystal data. Disturbances of the artificial data including Gaussian noise, stretching, and randomly missing particles are used to investigate the methods thoroughly. A popular method, called bond order parameter, has been repeatedly criticized as a structure analysis tool and will be improved with the help of a recent development. The method is called the bcc-sensitive Minkowski structure metric. It enhances robustness and consistency, while remaining compatible with previous bond-order-based results. Also, a promising method rooted in the molecular dynamics community is tested, yielding detailed insight of bond-order-specific drawbacks. With this investigation, the state of three-dimensional plasma crystal analysis will be significantly improved.
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Molecular monitoring of chronic myeloid leukemia patients using robust BCR-ABL1 tests standardized to the International Scale (IS) is key to proper disease management, especially when treatment cessation is considered. Most laboratories currently use a time-consuming sample exchange process with reference laboratories for IS calibration. A World Health Organization (WHO) BCR-ABL1 reference panel was developed (MR(1)-MR(4)), but access to the material is limited. In this study, we describe the development of the first cell-based secondary reference panel that is traceable to and faithfully replicates the WHO panel, with an additional MR(4.5) level. The secondary panel was calibrated to IS using digital PCR with ABL1, BCR and GUSB as reference genes and evaluated by 44 laboratories worldwide. Interestingly, we found that >40% of BCR-ABL1 assays showed signs of inadequate optimization such as poor linearity and suboptimal PCR efficiency. Nonetheless, when optimized sample inputs were used, >60% demonstrated satisfactory IS accuracy, precision and/or MR(4.5) sensitivity, and 58% obtained IS conversion factors from the secondary reference concordant with their current values. Correlation analysis indicated no significant alterations in %BCR-ABL1 results caused by different assay configurations. More assays achieved good precision and/or sensitivity than IS accuracy, indicating the need for better IS calibration mechanisms.
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Proteínas de Fusão bcr-abl/análise , Calibragem , Proteínas de Fusão bcr-abl/normas , Genes abl , Humanos , Reação em Cadeia da Polimerase , Proteínas Proto-Oncogênicas c-bcr/genética , Padrões de Referência , Organização Mundial da SaúdeRESUMO
While the initial treatment for primary hyperparathyroidism (pHPT), if managed by an experienced surgeon, is almost always successful, reoperations are challenging. Patients are at high risk for complications and the rates of success are plainly below those of primary cervical explorations. In this paper the reasons for failure during initial procedures are reviewed, as are the most important localization procedures and the prerequisites with regard to technical infrastructure as well as personnel, when planning repeat operations for a missed parathyroid adenoma. Provided that a standardized diagnostic and surgical approach is used, the surgeon is experienced, and up-to-date technical equipment is available, permanent normocalcemia following reoperations in pHPT is more frequently achieved than it used to be. The best option to avoid reoperations and associated complications is a successful initial intervention by an experienced surgeon. However, reoperations should always be performed by an experienced surgeon.
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Adenoma/cirurgia , Hiperparatireoidismo/cirurgia , Recidiva Local de Neoplasia/cirurgia , Neoplasias das Paratireoides/cirurgia , Adenoma/sangue , Adenoma/diagnóstico , Cálcio/sangue , Diagnóstico Diferencial , Humanos , Hiperparatireoidismo/sangue , Hiperparatireoidismo/diagnóstico , Recidiva Local de Neoplasia/sangue , Neoplasias Primárias Múltiplas/sangue , Neoplasias Primárias Múltiplas/diagnóstico , Neoplasias Primárias Múltiplas/cirurgia , Hormônio Paratireóideo/sangue , Neoplasias das Paratireoides/sangue , Neoplasias das Paratireoides/diagnóstico , Paratireoidectomia/métodos , Complicações Pós-Operatórias/etiologiaRESUMO
Tyrosine kinase inhibitors (TKI) have changed the natural course of chronic myeloid leukemia (CML). With the advent of second-generation TKI safety and efficacy issues have gained interest. The randomized CMLâ-âStudy IV was used for a long-term evaluation of imatinib (IM). 1503 patients have received IM, 1379 IM monotherapy. After a median observation of 7.1 years, 965 patients (64%) still received IM. At 10 years, progression-free survival was 82%, overall survival 84%, 59% achieved MR(5), 72% MR(4.5), 81% MR(4), 89% major molecular remission and 92% MR(2) (molecular equivalent to complete cytogenetic remission). All response levels were reached faster with IM800 mg except MR(5). Eight-year probabilities of adverse drug reactions (ADR) were 76%, of grades 3-4 22%, of non-hematologic 73%, and of hematologic 28%. More ADR were observed with IM800 mg and IM400 mg plus interferon α (IFN). Most patients had their first ADR early with decreasing frequency later on. No new late toxicity was observed. ADR to IM are frequent, but mostly mild and manageable, also with IM 800 mg and IM 400 mg+IFN. The deep molecular response rates indicate that most patients are candidates for IM discontinuation. After 10 years, IM continues to be an excellent initial choice for most patients with CML.
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Antineoplásicos/administração & dosagem , Benzamidas/administração & dosagem , Leucemia Mielogênica Crônica BCR-ABL Positiva/tratamento farmacológico , Piperazinas/administração & dosagem , Pirimidinas/administração & dosagem , Idoso , Antineoplásicos/efeitos adversos , Benzamidas/efeitos adversos , Progressão da Doença , Intervalo Livre de Doença , Feminino , Seguimentos , Proteínas de Fusão bcr-abl/genética , Proteínas de Fusão bcr-abl/metabolismo , Humanos , Mesilato de Imatinib , Interferon-alfa/uso terapêutico , Masculino , Pessoa de Meia-Idade , Projetos Piloto , Piperazinas/efeitos adversos , Inibidores de Proteínas Quinases/administração & dosagem , Inibidores de Proteínas Quinases/efeitos adversos , Pirimidinas/efeitos adversos , Indução de Remissão , Resultado do TratamentoRESUMO
Studies with idazoxan (IDA), a specific alpha 2-noradrenergic receptor antagonist, demonstrate effects on feeding behavior opposite to those observed with norepinephrine in the paraventricular nucleus (PVN) and peripheral injection of the alpha 2 agonist clonidine. Administration of IDA, both intraperitoneally (IP) and into the PVN at the onset of the nocturnal feeding cycle, caused a dose-related, selective suppression of carbohydrate intake 90 min after injection. To characterize further the impact of this antagonist on macronutrient intake, we examined in IDA-injected animals the macrostructure of feeding using computer-assisted analyses of meal patterns. Both IP and PVN administration of IDA produced a selective suppression of carbohydrate intake, primarily during the first meal of the feeding cycle. This effect occurred through significant reductions in meal size, diet composition, feeding time, and feeding rate for this nutrient. Idazoxan administration into the PVN continued to decrease carbohydrate intake in the next two meals and reduced the satiating impact of this nutrient. In contrast to this immediate change in carbohydrate intake, PVN IDA reduced protein intake after a latency of 4 h. although fat intake was suppressed only after a latency of 7 h. An increase in total meal number and a decrease in the average meal size across the 12-h dark cycle were seen after PVN IDA administration. These results, showing effects of peripheral and PVN-injected IDA on carbohydrate intake, suggest a possible physiological role of endogenous PVN alpha 2-noradrenergic receptors in modulating natural patterns of carbohydrate feeding at the onset of the dark period.
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Antagonistas Adrenérgicos alfa/farmacologia , Carboidratos da Dieta/administração & dosagem , Dioxanos/farmacologia , Comportamento Alimentar/efeitos dos fármacos , Preferências Alimentares/efeitos dos fármacos , Núcleo Hipotalâmico Paraventricular/efeitos dos fármacos , Receptores Adrenérgicos/efeitos dos fármacos , Animais , Gorduras na Dieta/administração & dosagem , Proteínas Alimentares/administração & dosagem , Relação Dose-Resposta a Droga , Ingestão de Energia/efeitos dos fármacos , Ingestão de Energia/fisiologia , Comportamento Alimentar/fisiologia , Preferências Alimentares/fisiologia , Idazoxano , Masculino , Núcleo Hipotalâmico Paraventricular/fisiologia , Ratos , Ratos Sprague-Dawley , Receptores Adrenérgicos/fisiologiaRESUMO
OBJECTIVE: To assess the level of a cohort of pediatric otolaryngologists' knowledge and understanding of genetics and genetic testing for deafness and hard of hearing (D/HOH). METHODS: A questionnaire was designed to assess the level of knowledge and understanding of the genetic basis and genetic testing for deafness among a cohort of pediatric otolaryngologists. Three hundred questionnaires were made available to attendees of the 14th (1999) Annual Meeting of the American Society of Pediatric Otolaryngology, Palm Desert, Calif. A series of questions asked to gauge the respondent's level of knowledge of genetics and hearing impairment addressed estimating recurrence risks for deaf and normal-hearing parents and the likelihood of detecting a mutation in connexin 26 in specific clinical scenarios. RESULTS: A total of 28 questionnaires were completed and returned. All respondents reported that they regularly saw patients for D/HOH. Almost half commonly refer these patients for genetic testing and counseling. Seventeen (71%) of 24 otolaryngologists stated they offered genetic testing in all situations, while 6 offered counseling only at parental request or to address recurrence risk issues. One otolaryngologist offered genetic testing if there was a deaf sibling. Twelve (67%) of 18 offered pretest counseling, which was most frequently provided by a genetic counselor. Although 3 (19%) of 16 otolaryngologists provided the counseling themselves, 2 (13%) reported that they and a genetic counselor provided the counseling. While 24 (89%) of the 27 correctly stated that nonsyndromic D/HOH is usually autosomal recessive, recurrence risks were incorrectly estimated in several examples. CONCLUSIONS: While the surveyed pediatric otolaryngologists have a good knowledge of genetics and genetic testing for D/HOH, recurrence risks were often inaccurate. Since D/HOH testing is clinically available, it is imperative that physicians are educated about genetics and genetic testing and are able to communicate this to their patients and their patients' families.
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Competência Clínica , Surdez/genética , Testes Genéticos , Otolaringologia , Pediatria , Criança , Coleta de Dados , Surdez/diagnóstico , Escolaridade , Aconselhamento Genético , Humanos , Projetos Piloto , Encaminhamento e Consulta , Fatores de RiscoRESUMO
RATIONALE AND OBJECTIVES: The purpose of this study was to examine the effects of placing a metal stent across a bronchial orifice. MATERIALS AND METHODS: Nine pigs were used as test subjects, because the right upper lobe bronchus comes directly off the trachea in these animals. One of three types of metal stents was placed into the trachea of each pig and covered the orifice of the right upper lobe bronchus. Follow-up studies were performed at 1 and 3 months to evaluate the right upper lobe for signs of bronchial obstruction, infection, and atelectasis. The animals were sacrificed at 3 months to study the histopathologic changes of the trachea and lungs. RESULTS: Two upper lobe bronchi remained patent; seven were obstructed by granulation tissue or plugs of mucus and inflammatory cells. Right upper lobe infiltration and atelectasis were seen in eight animals. Interestingly, radiographic opacities were also common in other lung segments. There was a tendency toward fewer and less extensive lung opacities at 3 months compared with that at 1 month. At histopathologic examination, areas of both acute and chronic pneumonia were found in the right upper lobe of all animals. The segment of trachea covered by the stent was lined with a thin layer of granulation tissue containing neutrophils, monocytes, and lymphocytes. The stent luminal surface was covered with columnar, cuboidal, and stratified squamous epithelium. Tracheal stenosis was seen in three animals because of excessive granulation tissue in two and a collapsed stent in one. CONCLUSION: Placement of metal stents in pig trachea covering the orifice of the right upper lobe bronchus resulted in retention of secretions and secondary infection in the right upper lobe and other distant lung segments.
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Brônquios/patologia , Modelos Animais de Doenças , Stents , Traqueia/patologia , Animais , Broncografia , Suínos , Traqueia/diagnóstico por imagemRESUMO
We characterize the specificity of a polyclonal antibody against heat shock protein 60 (hsp60) and present an application for ultrastructural localization studies of this protein. The antibody was obtained from an IgG fraction (AB 121) originally raised against the calcium binding protein calsequestrin by immunoabsorption on isolated rat liver hsp60. As shown by partial N-terminal amino acid sequence analysis of immunoprecipitated proteins AB 121 contained reactivities against hsp60, calsequestrin and the glycoprotein fetuin. In rat heart AB 121 recognized calsequestrin and hsp60. In human and rat liver the only reacting protein was hsp60. In rat erythrocytes the antibody bound to 61 kDa and 58 kDa isoforms of fetuin. According to published data no amino acid sequence homologies nor common motifs are found between calsequestrin, hsp60 and fetuin. As the first application the anti-hsp60 antibody was used for immuno-gold electron microscopical localization of hsp60: in myocardiocytes and hepatocytes of the rat strong labelling was obtained exclusively in mitochondria. No extramitochondrial structures were labelled. The specificity of the antibody and its ability to be visualized by immuno-gold electron microscopy offers the possibility to study the expression of this protein in the liver and in other organs. Possible clinical applications of these studies are discussed, since hsp60 could be a target antigen of autoantibodies in diseases such as autoimmune hepatitis, primary sclerosing cholangitis or primary biliary cirrhosis.