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1.
Prev Vet Med ; 218: 105999, 2023 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-37619426

RESUMO

Paratuberculosis (Johne´s disease) is a world-wide cattle disease caused by Mycobacterium avium subsp. paratuberculosis (MAP), associated with substantial economic losses. Purchase of subclinically infected animals or contact with animals and equipment of infected farms are known risk factors for disease transmission among herds. The aim of the present study was to identify specific management factors in Austria that triggered a MAP-positive herd status and to evaluate known risk factors for the transmission in cattle in small structured alpine agricultural systems. The agriculture in the Austrian province of Tyrol is characterized by smallholder structures, including shared alpine pastures and traditional barn management techniques. The data from an extensive survey with 50 questions in 2013/2014 and the development of the MAP herd status of 5592 cattle farms by taking feces and blood samples were examined and statistically evaluated. MAP herd status was determined by combining the results of boot swab samples, manure samples, pooled and individual feces samples as well as serological antibody testing by ELISA. The statistical analysis (odds ratio; OR) showed that the use of milk replacers for calf feeding (p = 0.047, OR=0.472) and the use of straw as bedding material for cows (p = 0.032, OR=0.625) were associated with a decreased chance of being a MAP-positive herd. Further, housing cows in deep litter systems (p = 0.028, OR=2.232), the presence of slurry channels (p = 0.028, OR=1.411) and the use of solid manure in young cattle (p = 0.041, OR=1.744) were associated with an increased OR for being MAP-positive. Surprisingly, sharing of lowland pastures (p = 0.564, OR=1.080), alpine pastures (p = 0.419, OR=1.143) or farm equipment (p = 0.733, OR=0.963) and farm size (p = 0.425) had no significant influence on the MAP herd status. The identified differences compared with previously published results in respect of MAP spread in cattle might be attributed to the traditional agricultural structures, including small family-based farms and common pasture during summer in alpine regions. Results of this study contribute to the understanding of the spread of MAP in cattle farming in alpine regions.

2.
Schweiz Arch Tierheilkd ; 161(10): 659-665, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31586928

RESUMO

INTRODUCTION: Subclinical mastitis is a frequent and economically important disease in cattle. Since the milk appears macroscopically normal the diagnosis depends on indirect parameters. In automatic milking systems ideally the measurements have to be automatized and completed during milking process. Oxygen concentration of the milk is a parameter which has not been used widely to date. Oxygen is consumed by cells in the milk, hence an increased number of cells results in decreased oxygen concentration. The main objectives of the study were to study the association of milk oxygen concentration with the number of cells and the electric conductivity and to assess the feasibility of oxygen concentration for detection of subclinical mastitis. The study was performed on five dairy farms using 690 macroscopically normal quarter milk samples. Oxygen concentrations (OC), somatic cell count (SCC) and electric conductivity (EC) were measured. The associations between the parameters were calculated and the diagnostic value of OC for detection of subclinical mastitis was estimated. Significant correlations were found between OC and SCC (rs =-0.28) and between OC and EC (rs = -0.20). SCC of the samples varied between 1,000 and 21,602,000 cells/ml. Out of the 690 milk samples 404 had SCC 200,000 cells/ml (subclinical mastitis). OC decreased whereas EC increased significantly with increasing SCC (P < 0.001). The threshold for OC to diagnose subclinical mastitis was 5.22% (sensitivity 84%, specificity 46%). The area under the curve describing the diagnostic value of OC was 0.72 (confidence interval 0.68-0.76). In conclusion OC has potential to be used as parameter to detect subclinical mastitis in dairy cattle. Currently the application is not suitable for routine use. Further research is necessary to improve measurement technology and diagnostic value of the parameter.


INTRODUCTION: La mammite subclinique est une maladie fréquente et économiquement importante chez les bovins. Puisque le lait apparaît macroscopiquement normal, le diagnostic dépend des paramètres indirects. Dans les systèmes de traite automatique, les mesures doivent idéalement être automatisées et effectuées pendant le processus de traite. La concentration en oxygène du lait est un paramètre qui n'a pas été largement utilisé à ce jour. L'oxygène est consommé par les cellules dans le lait, un nombre accru de cellules entraînant donc une diminution de la concentration en oxygène. Les principaux objectifs de l'étude étaient d'étudier l'association de la concentration d'oxygène du lait avec le nombre de cellules et la conductivité électrique et d'évaluer la praticabilité de la mesure de la concentration en oxygène pour la détection de la mammite subclinique. L'étude a été réalisée dans cinq fermes laitières à l'aide de 690 échantillons de lait de quartier macroscopiquement normaux. Des concentrations d'oxygène (OC), le nombre de cellules somatiques (SCC) et la conductivité électrique (EC) ont été mesurés. Les associations entre les paramètres ont été calculées et la valeur diagnostique de l'OC pour la détection de la mammite subclinique a été estimée. Des corrélations significatives ont été trouvées entre OC et SCC (rs= -0,28) et entre OC et EC (rs = -0,20). Le SCC des échantillons variait entre 1'000 et 21'602'000 cellules/ml. Sur les 690 échantillons de lait, 404 avaient un SCC de 200 000 cellules/ml (mammite subclinique). L'OC a diminué alors que l'EC a augmenté de façon significative avec l'augmentation du SCC (P - 0,001). Le seuil pour que l'OC permette un diagnostic de mammite subclinique était de 5,22 % (sensibilité 84 %, spécificité 46 %). La zone sous la courbe décrivant la valeur diagnostique de CO était 0.72 (intervalle de confiance 0.68-0.76). En conclusion, l'OC pourrait être utilisée comme paramètre pour détecter la mammite subclinique chez les bovins laitiers. Actuellement, l'application n'est pas adaptée à une utilisation de routine. D'autres recherches sont nécessaires pour améliorer la technologie de mesure et la valeur diagnostique du paramètre.


Assuntos
Mastite Bovina/diagnóstico , Leite/química , Oxigênio/análise , Animais , Infecções Assintomáticas , Bovinos , Indústria de Laticínios , Condutividade Elétrica , Feminino , Leite/citologia , Leite/fisiologia , Curva ROC
3.
Insights Imaging ; 4(4): 481-90, 2013 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-23673453

RESUMO

OBJECTIVES: The purpose of this study was to evaluate if positron emission tomography (PET)/magnetic resonance imaging (MRI) with just one gradient echo sequence using the body coil is diagnostically sufficient compared with a standard, low-dose non-contrast-enhanced PET/computed tomography (CT) concerning overall diagnostic accuracy, lesion detectability, size and conspicuity evaluation. METHODS AND MATERIALS: Sixty-three patients (mean age 58 years, range 19-86 years; 23 women, 40 men) referred for either staging or restaging/follow-up of various malignant tumours (malignant melanoma, lung cancer, breast cancer, Hodgkin's lymphoma, non-Hodgkin's lymphoma, CUP, gynaecology tumours, pleural mesothelioma, oesophageal cancer, colorectal cancer, stomach cancer) were prospectively included. Imaging was conducted using a tri-modality PET/CT-MR set-up (full ring, time-of-flight Discovery PET/CT 690, 3 T Discovery MR 750, both GE Healthcare, Waukesha, WI). All patients were positioned on a dedicated PET/CT- and MR-compatible examination table, allowing for patient transport from the MR system to the PET/CT without patient movement. In accordance with RECIST 1.1 criteria, measurements of the maximum lesion diameters on CT and MR images were obtained. In lymph nodes, the short axis was measured. A four-point scale was used for assessment of lesion conspicuity: 1 (>25 % of lesion borders definable), 2 (25-50 %), 3 (50-75 %) and 4 (>75 %). For each lesion the corresponding anatomical structure was noted based on anatomical information of the spatially co-registered PET/CT and PET/MRI image sections. Additionally, lesions were divided into three categories: "tumour mass", "lymph nodes" and "lesions". Differences in overall lesion detectability and conspicuity in PET/CT and PET/MRI, as well as differences in detectability based on the localisation and lesion type, were analysed by Wilcoxon signed rank test. RESULTS: A total of 126 PET-positive lesions were evaluated. Overall, no statistically significant superiority of PET/CT over PET/MRI or vice versa in terms of lesion conspicuity was found (p = 0.095; mean score CT 2.93, mean score MRI 2.75). A statistically significant superiority concerning conspicuity of PET/CT over PET/MRI was found in pulmonary lesions (p = 0.016). Additionally, a statistically significant superiority of PET/CT over PET/MRI in "lymph nodes" regarding lesion conspicuity was also found (p = 0.033). A higher mean score concerning bone lesions were found for PET/CT compared with PET/MRI; however, these differences did not achieve statistical significance. CONCLUSION: Overall, PET/MRI with body coil acquisition does not match entirely the diagnostic accuracy of standard low-dose PET/CT. Thus, it might only serve as a back-up solution in very few patients. Overall, more time needs to be invested on the MR imaging part (higher matrix, more breath-holds, additional surface coil acquired sequences) to match up with the standard low-dose PET/CT. MAIN MESSAGES: • Evaluation of whether PET/MRI with one sequence using body coil is diagnostically sufficient compared with PET/CT • PET/MRI with body coil does not match entirely the diagnostic accuracy of standard low-dose PET/CT • PET/MRI might only serve as a backup solution in patients.

6.
J Anim Sci ; 87(4): 1548-57, 2009 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-18952722

RESUMO

Ninety-three crossbred steer calves (BW+/-SD=385+/-50 kg) were used (n=48 steers in yr 1, n=45 steers in yr 2) to examine the relationship among carcass traits, lean, bone, and fat proportions, visceral tissue weights, and pancreatic digestive enzyme activity with DMI, ADG, G:F, and residual feed intake. Calves were progeny from crossbred dams predominantly of Angus and Simmental breeding and were sired by Angus, Simmental, crossbred (predominantly of Angus and Simmental breeding), Charolais, or Piedmontese bulls. Steers were fed a high-moisture corn-based diet for an average of 112 d. Partial correlation analysis accounting for year, pen within year, week of slaughter within year, and sire breed was conducted. Gain:feed was negatively correlated (P 0.10) between performance measures and the pancreatic proportional content of alpha-amylase and trypsin activity (units/kg of BW). These data indicate that carcass fatness traits and changes in the proportional weight of total viscera may be negatively associated with G:F and that visceral fat weight proportion and trim and kidney fat weight proportion may be important factors influencing this relationship.


Assuntos
Composição Corporal/fisiologia , Bovinos/crescimento & desenvolvimento , Bovinos/metabolismo , Pâncreas/enzimologia , Tecido Adiposo/metabolismo , Animais , Ingestão de Alimentos/fisiologia , Masculino , Tamanho do Órgão/fisiologia , Vísceras/anatomia & histologia , Aumento de Peso/fisiologia
7.
BMC Bioinformatics ; 8: 156, 2007 May 16.
Artigo em Inglês | MEDLINE | ID: mdl-17506883

RESUMO

BACKGROUND: The combination of mass spectrometry and solution phase amide hydrogen/deuterium exchange (H/D exchange) experiments is an effective method for characterizing protein dynamics, and protein-protein or protein-ligand interactions. Despite methodological advancements and improvements in instrumentation and automation, data analysis and display remains a tedious process. The factors that contribute to this bottleneck are the large number of data points produced in a typical experiment, each requiring manual curation and validation, and then calculation of the level of backbone amide exchange. Tools have become available that address some of these issues, but lack sufficient integration, functionality, and accessibility required to address the needs of the H/D exchange community. To date there is no software for the analysis of H/D exchange data that comprehensively addresses these issues. RESULTS: We have developed an integrated software system for the automated analysis and representation of H/D exchange data that has been titled "The Deuterator". Novel approaches have been implemented that enable high throughput analysis, automated determination of deuterium incorporation, and deconvolution of overlapping peptides. This has been achieved by using methods involving iterative theoretical envelope fitting, and consideration of peak data within expected m/z ranges. Existing common file formats have been leveraged to allow compatibility with the output from the myriad of MS instrument platforms and peptide sequence database search engines.A web-based interface is used to integrate the components of The Deuterator that are able to analyze and present mass spectral data from instruments with varying resolving powers. The results, if necessary, can then be confirmed, adjusted, re-calculated and saved. Additional tools synchronize the curated calculation parameters with replicate time points, increasing throughput. Saved results can then be used to plot deuterium buildup curves and 3D structural overlays. The system has been used successfully in a production environment for over one year and is freely available as a web tool at the project home page http://deuterator.florida.scripps.edu. CONCLUSION: The automated calculation and presentation of H/D exchange data in a user interface enables scientists to organize and analyze data efficiently. Integration of the different components of The Deuterator coupled with the flexibility of common data file formats allow this system to be accessible to the broadening H/D exchange community.


Assuntos
Medição da Troca de Deutério/métodos , Deutério/química , Hidrogênio/química , Software , Amidas/química , Sequência de Aminoácidos , Medição da Troca de Deutério/estatística & dados numéricos , Espectrometria de Massas/métodos , Espectrometria de Massas/estatística & dados numéricos , Dados de Sequência Molecular , Fragmentos de Peptídeos/química , Fragmentos de Peptídeos/genética , Software/estatística & dados numéricos
8.
Biochim Biophys Acta ; 1463(1): 142-50, 2000 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-10631303

RESUMO

In the present study, unilamellar liposomes coated with the crystalline bacterial cell surface layer (S-layer) protein of Bacillus stearothermophilus PV72/p2 were used as matrix for defined binding of functional molecules via the avidin- or streptavidin-biotin bridge. The liposomes were composed of dipalmitoyl phosphatidylcholine, cholesterol and hexadecylamine in a molar ratio of 10:5:4 and they had an average size of 180 nm. For introducing specific functions into the S-layer lattice without affecting substances encapsulated within the liposomes, crosslinking and activation reagents had to be identified which did not penetrate the liposomal membrane. Among different reagents, a hydrophilic dialdehyde generated by periodate cleavage of raffinose and a sulfo-succinimide activated dicarboxylic acid were found to be impermeable for the liposomal membrane. Both reagents completely crosslinked the S-layer lattice without interfering with its regular structure. Biotinylation of S-layer-coated liposomes was achieved by coupling p-diazobenzoyl biocytin which preferably reacts with the phenolic residue of tyrosine or with the imidazole ring of histidine. By applying this method, two biotin residues accessible for subsequent avidin binding were introduced per S-layer subunit. As visualized by labeling with biotinylated ferritin, an ordered monomolecular layer of streptavidin was formed on the surface of the S-layer-coated liposomes. As a second model system, biotinylated anti-human IgG was attached via the streptavidin bridge to the biotinylated S-layer-coated liposomes. The biological activity of the bound anti-human IgG was confirmed by ELISA.


Assuntos
Proteínas de Bactérias/química , Proteínas de Bactérias/metabolismo , Lipossomos , Glicoproteínas de Membrana/química , Glicoproteínas de Membrana/metabolismo , Aminas , Anticorpos Anti-Idiotípicos/metabolismo , Biotina , Carbodi-Imidas , Reagentes de Ligações Cruzadas , Cristalização , Humanos , Hidrocarbonetos , Imunoglobulina G/metabolismo , Técnicas In Vitro , Microscopia Eletrônica , Ligação Proteica
9.
Euro Surveill ; 5(11): 123-126, 2000 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-12631963

RESUMO

Sixteen culture confirmed cases of enteric infection with Salmonella enterica serotype Oranienburg were detected between August 10 and September 29 1999 in Tyrol (Austria). Ten of them suffered bloody diarrhoea and six were asymptomatic carriers. Intervie

10.
Biochim Biophys Acta ; 1418(1): 106-16, 1999 Apr 14.
Artigo em Inglês | MEDLINE | ID: mdl-10209215

RESUMO

Isolated subunits of the crystalline cell surface layer (S-layer) protein of Bacillus stearothermophilus PV72/p2 were recrystallized on positively charged unilamellar liposomes. Liposomes were composed of dipalmitoylphosphatidylcholine (DPPC), cholesterol and hexadecylamine (HDA) in a molar ratio of 10:5:4 and they were prepared by the dehydration-rehydration method followed by an extrusion procedure. The S-layer protein to DPPC ratio was 5.7 nmol/micromol which approximately corresponds to the theoretical value estimated by using the areas occupied by the S-layer lattice and the lipid membrane. Coating of the positively charged liposomes with S-layer protein resulted in inversion of the zeta-potential from +29.1 mV to -27.1 mV. Covalent crosslinking of the recrystallized S-layer protein was achieved with glutaraldehyde. Chemical analysis revealed that almost all amino groups (>95%) from HDA in the liposomal membrane were involved in the reaction. To study the influence of an S-layer lattice on the stability of the liposomes, the hydrophilic marker carboxyfluoresceine (CF) was encapsulated and its release was determined for plain and S-layer-coated liposomes in the course of mechanical and thermal challenges. In comparison to plain liposomes, S-layer-coated liposomes released only half the amount of enclosed CF upon exposure to shear forces or ultrasonication as mechanical stress factors. Furthermore, temperature shifts from 25 degrees C to 55 degrees C and vice versa induced considerably less CF release from S-layer-coated than from plain liposomes. A similar stabilizing effect of the S-layer lattice was observed after glutaraldehyde treatment of plain and S-layer-coated liposomes.


Assuntos
Proteínas de Bactérias/química , Geobacillus stearothermophilus/química , Lipossomos/química , Proteínas de Membrana/química , Cristalização , Fluoresceínas , Geobacillus stearothermophilus/genética , Glutaral , Lipossomos/ultraestrutura , Estresse Mecânico , Temperatura , Ultrassom
11.
Wien Med Wochenschr ; 151(5-6): 126-30, 1999.
Artigo em Alemão | MEDLINE | ID: mdl-11315410

RESUMO

Boyle's law is important for the understanding of the pathophysiology of barotrauma. Barotrauma--any disease or injury due to unequal pressures between a hollow space inside the body and the ambient pressure, or between two spaces within the body; examples include arterial gas embolism, pneumomediastinum, and pneumothorax. It may affect the lungs, the middle ear, the inner ear, the external ear, the eyes, the teeth, the gastrointestinal tract, the skin. Symptoms remain the same whether it all happens during ascent or descent.


Assuntos
Barotrauma/complicações , Mergulho/lesões , Barotrauma/fisiopatologia , Cavidades Cranianas/lesões , Sistema Digestório/lesões , Orelha/lesões , Orelha/fisiopatologia , Traumatismos Oculares/etiologia , Traumatismos Faciais/etiologia , Humanos , Seios Paranasais/lesões , Púrpura/etiologia , Síndrome do Desconforto Respiratório/etiologia , Traumatismos Dentários/etiologia
12.
Mol Membr Biol ; 15(2): 69-74, 1998.
Artigo em Inglês | MEDLINE | ID: mdl-9724924

RESUMO

Isolated S-layer subunits from Bacillus coagulans E38-66/v1 were recrystallized on positively charged, unilamellar liposomes composed of dipalmitoylphosphatidylcholine, cholesterol and hexadecylamine. The thermotropic phase behaviour of S-layer coated and uncoated liposomes was characterized by differential scanning microcalorimetry indicating for both preparations a broad transition around 50 degrees C due to the chain-melting from a liquid-ordered gel-like to a liquid-ordered fluid phase as described for phosphatidylcholine/cholesterol mixtures. The slightly higher phase transition temperature for the S-layer coated liposomes were explained by increased intermolecular order. Cross-linking the S-layer subunits covalently to hexadecylamine with glutaraldehyde induced phase separation within the liposomes. Based on deconvolution of the normalized excess heat capacity functions it was proposed that the different lipid domains arise from phospholipids representing different degrees of mobility.


Assuntos
Bacillus/química , Proteínas da Membrana Bacteriana Externa/química , Lipossomos/química , Calorimetria , Membrana Celular/química , Reagentes de Ligações Cruzadas , Conformação Proteica
13.
J Oral Rehabil ; 25(7): 513-26, 1998 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-9722098

RESUMO

The survival rate and the reasons for failures of 130 combined fixed-removable reconstructions (CFR), incorporated in 112 patients, were assessed. Each CFR reconstruction was classified depending on its attachments: 76 reconstructions were attached with rigid, precise attachments, and constituted the rigid group; 54 reconstructions were attached with either semi-precision or individual attachments and were defined as the semi-rigid group. Of the 130 reconstructions, 41 were determined as complete successes, 39 as partial successes and 50 as failures, leading to 37 major repairs and to 13 new reconstructions. Three reconstructions failed due to technical reasons, 36 due to biological reasons and for 11 reconstructions, both categories of reasons were responsible for their failure. In total, technical reasons were counted 15 times in comparison to 73 biological reasons for those 50 failed reconstruction, with 29 fractured abutment teeth as the most common biological reason. Within the rigid group, 45 failed reconstructions were observed, whereas within the semi rigid group only 5 failures occurred, leading to an 8-year survival estimate (+/- SD) of 30.1% (+/- 6.9%) for the rigid group and 93.1% (+/- 3.9%) for the semi rigid group. Beside the attachment type, the anatomy of the partially edentulous tooth arch in form of the free-end situation and the dentate opposing jaw were identified as risk factors.


Assuntos
Falha de Restauração Dentária , Retenção de Dentadura/instrumentação , Prótese Parcial Fixa , Prótese Parcial Removível , Adulto , Idoso , Idoso de 80 Anos ou mais , Grampos Dentários , Planejamento de Dentadura , Encaixe de Precisão de Dentadura , Análise de Falha de Equipamento , Feminino , Humanos , Arcada Parcialmente Edêntula/patologia , Masculino , Pessoa de Meia-Idade , Modelos de Riscos Proporcionais , Análise de Regressão , Estudos Retrospectivos , Fatores de Risco , Análise de Sobrevida
14.
Allergy ; 53(8): 786-93, 1998 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-9722228

RESUMO

The incidence of allergy to airborne proteins derived from tree and grass pollen, feces of mites, spores of molds, and pet dander has been increasing over the last decades. Since precise diagnosis is a prerequisite for successful immunotherapy, there is a rising demand for rapid, reliable, and inexpensive screening methods such as dipstick assays. With the purified recombinant major birch-pollen allergen rBet v 1a as model protein, crystalline bacterial cell-surface layers (S-layers) were tested for their applicability as an immobilization matrix for dipstick development. For this purpose, S-layers were deposited on a mechanically stable microporous support, cross-linked with glutaraldehyde, and free carboxylic acid groups of the S-layer protein were activated with carbodiimide. In the present test system, rBet v 1a was immobilized via the monoclonal mouse antibody BIP 1, which, unlike the allergen, is too large to enter the pores of the S-layer lattice, and which therefore formed a closed monolayer on the outermost surface of the crystal lattice. Moreover, BIP 1 is known to modulate IgE binding to the allergen. After incubation of the dipsticks in serum, washing of the reaction zone under tap water, and binding of an anti-IgE alkaline phosphatase conjugate, 5-bromo-4-chloro-3-indolyl phosphate/nitro blue tetrazolium was used as substrate, forming an IgE concentration-dependent colored precipitate on the S-layer surface. The investigation of patient sera previously tested with the CAP system confirmed the specificity of the S-layer-based dipstick assay. Since the dipstick is easy to handle and the whole test procedure takes only 90 min, this test system should be applicable for rapid determination of specific IgE and for first screening in the doctor's practice.


Assuntos
Alérgenos/imunologia , Imunoensaio/métodos , Imunoglobulina E/sangue , Glicoproteínas de Membrana , Proteínas de Plantas/imunologia , Animais , Anticorpos Monoclonais/imunologia , Anticorpos Monoclonais/isolamento & purificação , Antígenos de Plantas , Bacillus anthracis/ultraestrutura , Proteínas de Bactérias , Parede Celular/ultraestrutura , Cristalização , Humanos , Imunoglobulina E/imunologia , Imunoadsorventes , Camundongos , Microscopia Eletrônica , Testes de Precipitina , Teste de Radioalergoadsorção , Proteínas Recombinantes/imunologia , Fatores de Tempo
15.
FEMS Microbiol Rev ; 20(1-2): 151-75, 1997 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-9276930

RESUMO

The wealth of information existing on the general principle of S-layers has revealed a broad application potential. The most relevant features exploited in applied S-layer research are: (i) pores passing through S-layers show identical size and morphology and are in the range of ultrafiltration membranes; (ii) functional groups on the surface and in the pores are aligned in well-defined positions and orientations and accessible for binding functional molecules in very precise fashion; (iii) isolated S-layer subunits from many organisms are capable of recrystallizing as closed monolayers onto solid supports at the air-water interface, on lipid monolayers or onto the surface of liposomes. Particularly their repetitive physicochemical properties down to the subnanometer scale make S-layers unique structures for functionalization of surfaces and interfaces down to the ultimate resolution limit. The following review focuses on selected applications in biotechnology, diagnostics, vaccine development, biomimetic membranes, supramolecular engineering and nanotechnology. Despite progress in the characterization of S-layers and the exploitation of S-layers for the applications described in this chapter, it is clear that the field lags behind others (e.g. enzyme engineering) in applying recent advances in protein engineering. Genetic modification and targeted chemical modification would allow several possibilities including the manipulation of pore permeation properties, the introduction of switches to open and close the pores, and the covalent attachment to surfaces or other macromolecules through defined sites on the S-layer protein. The application of protein engineering to S-layers will require the development of straightforward expression systems, the development of simple assays for assembly and function that are suitable for the rapid screening of numerous mutants and the acquisition of structural information at atomic resolution. Attention should be given to these areas in the coming years.


Assuntos
Bactérias/ultraestrutura , Biotecnologia/métodos , Membrana Celular , Parede Celular , Bactérias/química , Proteínas da Membrana Bacteriana Externa/química , Proteínas da Membrana Bacteriana Externa/ultraestrutura , Membrana Celular/química , Membrana Celular/ultraestrutura , Parede Celular/química , Parede Celular/ultraestrutura , Fenômenos Químicos , Físico-Química , Cristalização , Desenho de Fármacos , Lipossomos , Substâncias Macromoleculares , Lipídeos de Membrana/química , Metalurgia/métodos , Ligação Proteica , Conformação Proteica , Ultrafiltração/instrumentação , Vacinas/química
16.
Genome Res ; 7(5): 422-33, 1997 May.
Artigo em Inglês | MEDLINE | ID: mdl-9149939

RESUMO

We have constructed a physical map of the human genome by using a panel of 83 whole genome radiation hybrids (the Stanford G3 panel) in conjunction with 10,478 sequence-tagged sites (STSs) derived from random genomic DNA sequences, previously mapped genetic markers, and expressed sequences. Of these STSs, 5049 are framework markers that fall into 1766 high-confidence bins. An additional 945 STSs are indistinguishable in their map location from one or more of the framework markers. These 5994 mapped STSs have an average spacing of 500 kb. An additional 4484 STSs are positioned with respect to the framework markers. Comparison of the orders of markers on this map with orders derived from independent meiotic and YAC STS-content maps indicates that the error rate in defining high-confidence bins is < 5%. Analysis of 322 random cDNAs indicates that the map covers the vast majority of the human genome. This STS-based radiation hybrid map of the human genome brings us one step closer to the goal of a physical map containing 30,000 unique ordered landmarks with an average marker spacing of 100 kb.


Assuntos
Mapeamento Cromossômico/métodos , Cromossomos Humanos/genética , Genoma Humano , Células Híbridas/efeitos da radiação , Sitios de Sequências Rotuladas , Animais , Linhagem Celular , Cromossomos Humanos/efeitos da radiação , Cricetinae , Diploide , Marcadores Genéticos , Humanos , Linfócitos/patologia , Masculino , Software
17.
Science ; 274(5287): 540-6, 1996 Oct 25.
Artigo em Inglês | MEDLINE | ID: mdl-8849440

RESUMO

The human genome is thought to harbor 50,000 to 100,000 genes, of which about half have been sampled to date in the form of expressed sequence tags. An international consortium was organized to develop and map gene-based sequence tagged site markers on a set of two radiation hybrid panels and a yeast artificial chromosome library. More than 16,000 human genes have been mapped relative to a framework map that contains about 1000 polymorphic genetic markers. The gene map unifies the existing genetic and physical maps with the nucleotide and protein sequence databases in a fashion that should speed the discovery of genes underlying inherited human disease. The integrated resource is available through a site on the World Wide Web at http://www.ncbi.nlm.nih.gov/SCIENCE96/.


Assuntos
Mapeamento Cromossômico , Genoma Humano , Projeto Genoma Humano , Sequência de Aminoácidos , Animais , Sequência de Bases , Linhagem Celular , Cromossomos Artificiais de Levedura , Redes de Comunicação de Computadores , DNA Complementar/genética , Bases de Dados Factuais , Expressão Gênica , Marcadores Genéticos , Humanos , Família Multigênica , RNA Mensageiro/genética , Homologia de Sequência do Ácido Nucleico , Sitios de Sequências Rotuladas
18.
Psychosom Med ; 58(3): 211-6, 1996.
Artigo em Inglês | MEDLINE | ID: mdl-8771619

RESUMO

There is little empirical research on the familial and parental correlates of alexithymia. A two-part study explored how the affective and cognitive characteristics of alexithymia are related to family dysfunction and maternal alexithymia. In Part I, 127 young adults were evaluated for alexithymia with the Toronto Alexithymia Scale-20 (TAS-20), for impaired imagination with the Scored Archetypal Test with Nine Elements, and for family dysfunction with the McMaster Family Assessment Device. In Part II, 80 of their mothers completed the TAS-20 about themselves, and maternal alexithymia characteristics were correlated with those of their offspring. In Part I, general family pathology was associated with alexithymia. In particular, difficulty identifying feelings was related to dysfunctional family affective involvement, externally oriented thinking was related to deficient family behavior control, and impaired imagination was related to inadequate family problem solving; these relationships were independent of general family pathology and subjects' positive and negative affect. In Part II, maternal alexithymia characteristics were correlated significantly with the offsprings', controlling for both respondents' positive and negative affect. These findings implicate disturbed family functioning and maternal alexithymia in the development of alexithymia characteristics in children.


Assuntos
Sintomas Afetivos/psicologia , Saúde da Família , Mães/psicologia , Adolescente , Adulto , Conscientização , Filho de Pais com Deficiência , Emoções Manifestas , Feminino , Humanos , Imaginação , Transmissão Vertical de Doenças Infecciosas , Masculino , Poder Familiar , Análise de Regressão , Estudos de Amostragem , Fatores Sexuais
19.
Arzneimittelforschung ; 42(9): 1098-100, 1992 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-1445475

RESUMO

It is shown that it is possible to characterize sustained release formulations in vitro using not only dissolution data but also an absorption model system. The mean dissolution time (MDT) has been shown to be a suitable parameter for evaluating sustained release formulations in vitro. t1/2 and mean residence time (MRT) have been shown to be convenient pharmacokinetic parameters for characterizing sustained release formulations. For comparing in vitro and in vivo results the quotients MDT normal/MDT retard and MRT normal/MRT retard do seem to be useful.


Assuntos
Verapamil/farmacocinética , Adulto , Preparações de Ação Retardada , Feminino , Meia-Vida , Humanos , Absorção Intestinal , Masculino , Pessoa de Meia-Idade , Modelos Biológicos , Solubilidade , Verapamil/administração & dosagem
20.
J Histochem Cytochem ; 39(1): 123-9, 1991 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-1701182

RESUMO

Meprin is a membrane-bound metalloproteinase which is expressed at high levels in the brush border membrane of proximal tubules of kidneys of some mouse strains (referred to as high meprin-activity mice). The mature active proteinase is not present in kidneys of many inbred strains of mice; however, these low meprin-activity mice possess a kidney protein that crossreacts with polyclonal antibodies prepared against meprin. In the present studies, immunohistochemical methods were used to determine the presence of meprin in liver, pancreas, spleen, testis, thymus, kidney, salivary glands, stomach, duodenum, and skin. Meprin crossreactivity was observed only in kidney and salivary glands. In salivary glands, the enzyme was found on the luminal surface of intercalated and striated ducts of submandibular and parotid glands and on interlobular ducts of the latter. In both kidney and salivary glands, the intensity of immunochemical staining was greater in males compared with females. For both sexes, immunoreactivity was markedly greater in the high meprin-activity mice compared to the low meprin-activity mice. These studies indicate that meprin has a limited tissue distribution, and that genetic and hormonal factors that regulate the proteinase are similar in kidney and salivary glands. The localization of the proteinase implies that the enzyme functions in modifying proteins and peptides that are secreted or re-absorbed in the ducts of these tissues.


Assuntos
Técnicas Imunoenzimáticas , Glândulas Salivares/enzimologia , Tiopronina/análise , Animais , Feminino , Rim/enzimologia , Túbulos Renais Proximais/enzimologia , Masculino , Camundongos , Camundongos Endogâmicos C3H , Camundongos Endogâmicos C57BL , Microvilosidades/enzimologia , Glândula Parótida/enzimologia , Caracteres Sexuais , Coloração e Rotulagem , Glândula Submandibular/enzimologia , Distribuição Tecidual
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