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1.
Magn Reson Med ; 89(4): 1601-1616, 2023 04.
Artigo em Inglês | MEDLINE | ID: mdl-36478417

RESUMO

PURPOSE: Studies at 3T have shown that T1 relaxometry enables characterization of brain tissues at the single-subject level by comparing individual physical properties to a normative atlas. In this work, an atlas of normative T1 values at 7T is introduced with 0.6 mm isotropic resolution and its clinical potential is explored in comparison to 3T. METHODS: T1 maps were acquired in two separate healthy cohorts scanned at 3T and 7T. Using transfer learning, a template-based brain segmentation algorithm was adapted to ultra-high field imaging data. After segmenting brain tissues, volumes were normalized into a common space, and an atlas of normative T1 values was established by modeling the T1 inter-subject variability. A method for single-subject comparisons restricted to white matter and subcortical structures was developed by computing Z-scores. The comparison was applied to eight patients scanned at both field strengths for proof of concept. RESULTS: The proposed method for morphometry delivered segmentation masks without statistically significant differences from those derived with the original pipeline at 3T and achieved accurate segmentation at 7T. The established normative atlas allowed characterizing tissue alterations in single-subject comparisons at 7T, and showed greater anatomical details compared with 3T results. CONCLUSION: A high-resolution quantitative atlas with an adapted pipeline was introduced and validated. Several case studies on different clinical conditions showed the feasibility, potential and limitations of high-resolution single-subject comparisons based on quantitative MRI atlases. This method in conjunction with 7T higher resolution broadens the range of potential applications of quantitative MRI in clinical practice.


Assuntos
Imageamento por Ressonância Magnética , Substância Branca , Humanos , Imageamento por Ressonância Magnética/métodos , Substância Branca/diagnóstico por imagem , Algoritmos , Encéfalo/diagnóstico por imagem
2.
Hippocampus ; 31(12): 1257-1270, 2021 12.
Artigo em Inglês | MEDLINE | ID: mdl-34739173

RESUMO

The investigation of the physical traces of memories (engrams) has made significant progress in the last decade due to optogenetics and fluorescent cell tagging applied in rodents. Engram cells were identified. The ablation of engram cells led to the loss of the associated memory, silent memories were reactivated, and artificial memories were implanted in the brain. Human engram research lags behind engram research in rodents due to methodological and ethical constraints. However, advances in multivariate analysis techniques of functional magnetic resonance imaging (fMRI) data and machine learning algorithms allowed the identification of stable engram patterns in humans. In addition, MRI scanners with an ultrahigh field strength of 7 Tesla (T) have left their prototype state and became more common around the world to assist human engram research. Although most engram research in humans is still being performed with a field strength of 3T, fMRI at 7T will push engram research. Here, we summarize the current state and findings of human engram research and discuss the advantages and disadvantages of applying 7 versus 3T fMRI to image human memory traces.


Assuntos
Neurônios , Optogenética , Encéfalo/diagnóstico por imagem , Humanos , Imageamento por Ressonância Magnética
3.
Data Brief ; 41: 107877, 2022 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-35198664

RESUMO

Basin tests were performed at the Aalto Ice Tank to gather data on ice-structure action and interaction from ice failing against a vertically sided cylindrical pile. The tests were performed with a real-time hybrid test setup, which combined physical and numerical components to simulate a range of test structures in real-time. The dataset includes results from tests with offshore wind turbine structures, structural models representing a series of single- and multi-degree-of-freedom oscillators, and scaled dynamic models of the Norströmsgrund lighthouse and the Molikpaq caisson structure. In addition, forced vibration tests and rigid structure tests were performed. Ice loads and structural response were measured with accelerometers, displacement sensors, potentiometers, strain gauges and load cells and the ice-structure interaction process was filmed from three different camera angles. The resulting raw data have been categorized and stored as unfiltered time series. A total of 259 different tests are included in the dataset. The model ice formation procedure and the test temperature were aimed at creating model ice that mimics the material behavior of full-scale saline ice during crushing failure, with a specific focus on the transition from brittle to ductile behavior. The data can be used for validation of models for dynamic ice-structure interaction. The offshore wind turbine data can be used to study the effect of wind loading on the interaction with ice and the effect of the specific dynamic properties of wind turbine structures with monopile foundations on the ice-structure interaction process. The forced-oscillation data can be used to quantify the time and speed dependant aspects of ice loading. The Norströmsgrund lighthouse and the Molikpaq data can be used as a reference comparison to full-scale data on ice loads.

4.
Disabil Rehabil ; 42(14): 2076-2082, 2020 07.
Artigo em Inglês | MEDLINE | ID: mdl-30669878

RESUMO

Study design: This consisted of a translation and validation study.Background: Acute hamstring injury is a frequent muscle strain in sports that require high explosive strength, impulsion or running phases. Therefore, the Functional Assessment Scale for Hamstring Injury questionnaire was developed to assess pain, physical activity level and ability to perform various exercises in patients with hamstring injuries. The Functional Assessment Scale for Hamstring Injury questionnaire is currently available in English, German, and Greek.Objectives: The goal of this study was to provide a cross-culturally adapted French-translation of the FASH questionnaire and to assess its psychometric performance.Methods: The French-translation and cross-cultural adaptation process were based on international recommendations, following six rigorous steps: (a) two initial translations from English to French; (b) synthesis of the two translations; (c) back-translations; (d) comparisons between the back-translations and the original questionnaire by an expert committee; (e) pretest; and (f) approval of the final French version of the Functional Assessment Scale for Hamstring Injury questionnaire. To validate this French version, 116 subjects (17 pathological patients, 19 patients with other muscle injury, 40 athletes at risk, and 40 healthy control athletes) were recruited to complete the Functional Assessment Scale for Hamstring Injury questionnaire. The Short Form Health Survey (SF-36) was used as a comparative questionnaire. The psychometric properties of the questionnaire were evaluated by determining the test-retest reliability after a 48-60-h interval, internal consistency, construct validity, and floor/ceiling effects.Results: All of the items of the Functional Assessment Scale for Hamstring Injury questionnaire were translated without any major difficulties. The questionnaire showed excellent discriminative power by obtaining significantly different scores from the four groups (p = 0.01). Regarding psychometric performances, the test-retest reliability was excellent (IntraClass Coefficient Correlation of 0.997). Very high internal consistency was also observed (Cronbach's alpha of 0.969). Correlations with the physical health subscales of the SF-36 were significant and considered to be strong, indicating an excellent convergent validity. The other subscales of the SF-36 (mental health) were weakly correlated with the FASH, reflecting good divergent validity. No floor or ceiling effects were observed.Conclusion: The French-translation of the Functional Assessment Scale for Hamstring Injury questionnaire and its cross-cultural adaptation can be considered to be successful. Functional Assessment Scale for Hamstring Injury-French questionnaire is now a reliable and valid tool for patients suffering from acute hamstring injury, and its application in clinical practice is particularly relevant.Implications for rehabilitationThe FASH-F can be considered to be discriminant, reliable and valid for the evaluation of the severity of symptoms and sports ability in individuals with hamstring injuries.FASH-F is now a reliable and valid tool for French-speaking patients suffering from acute hamstring injury, and its application in clinical practice is particularly relevant.A limitation of our study could be that the distribution between the different study groups was not homogeneous implying that our findings may not be fully representative of the general population.


Assuntos
Atletas , Traumatismos em Atletas/diagnóstico , Músculos Isquiossurais/lesões , Inquéritos e Questionários/normas , Adulto , Traumatismos em Atletas/fisiopatologia , Comparação Transcultural , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Modalidades de Fisioterapia , Psicometria , Reprodutibilidade dos Testes , Tradução , Traduções
5.
J Virol Methods ; 276: 113786, 2020 02.
Artigo em Inglês | MEDLINE | ID: mdl-31765721

RESUMO

Foot-and-mouth disease virus (FMDV) is a highly variable RNA virus existing as seven different serotypes. The antigenic variability between and within serotypes can limit the cross-reactivity and therefore the in vivo cross-protection of vaccines. Selection of appropriate vaccine strains is crucial in the control of FMD. Determination of indirect relationships (r1-value) between potential vaccine strains and field strains based on antibody responses against both are routinely used for vaccine matching purposes. Aiming at the investigation of the repeatability, reproducibility and comparability of r1-value determination within and between laboratories and serological tests, a small scale vaccine matching ring test for FMDV serotype A was organized. Well-characterized serum pools from cattle vaccinated with a monovalent A24/Cruzeiro/Brazil/55 (A24) FMD vaccine with known in vivo protection status (homologous and heterologous) were distributed to four laboratories to determine r1-values for the heterologous FMD strains A81/Argentina/87, A/Argentina/2000 and A/Argentina/2001 using the virus neutralization tests (VNT) and liquid phase blocking ELISA (LPBE). Within laboratories, the repeatability of r1-value determination was high for both antibody assays. VNT resulted in reproducible and comparable r1-values between laboratories, indicative of a lack of antigenic relatedness between the A24 strain and the heterologous strains tested in this work, thus corresponding to some of the in vivo findings with these strains. Using LPBE, similar trends in r1-values were observed in all laboratories, but the overall reproducibility was lower than with VNT. Inconsistencies between laboratories may at least in part be attributed to differences in LPBE protocols as well as the in preexisting information generated in each laboratory (such as antibody titer-protection correlation curves). To gain more insight in the LPBE-derived r1-values standard bovine control sera were included in the antibody assays performed in each laboratory and a standardization exercise was performed.


Assuntos
Febre Aftosa/imunologia , Testes Sorológicos/normas , Testes Sorológicos/veterinária , Vacinas Virais/imunologia , Animais , Bovinos , Febre Aftosa/prevenção & controle , Testes de Neutralização , Variações Dependentes do Observador , Reprodutibilidade dos Testes , Vacinas Virais/administração & dosagem
6.
Elife ; 82019 07 22.
Artigo em Inglês | MEDLINE | ID: mdl-31329098

RESUMO

Social interactions have a major impact on well-being. While many individuals actively seek social situations, others avoid them, at great cost to their private and professional life. The neural mechanisms underlying individual differences in social approach or avoidance tendencies are poorly understood. Here we estimated people's subjective value of engaging in a social situation. In each trial, more or less socially anxious participants chose between an interaction with a human partner providing social feedback and a monetary amount. With increasing social anxiety, the subjective value of social engagement decreased; amygdala BOLD response during decision-making and when experiencing social feedback increased; ventral striatum BOLD response to positive social feedback decreased; and connectivity between these regions during decision-making increased. Amygdala response was negatively related to the subjective value of social engagement. These findings suggest a relation between trait social anxiety/social avoidance and activity in a subcortical network during social decision-making.


Assuntos
Tonsila do Cerebelo/fisiologia , Tomada de Decisões , Rede Nervosa/fisiologia , Assunção de Riscos , Comportamento Social , Estriado Ventral/fisiologia , Adulto , Feminino , Voluntários Saudáveis , Humanos , Masculino , Adulto Jovem
7.
Eur J Pharm Sci ; 34(4-5): 293-302, 2008 Aug 07.
Artigo em Inglês | MEDLINE | ID: mdl-18602800

RESUMO

Solid dispersion formulations made up of d-alpha-tocopheryl polyethylene glycol succinate 1000 (TPGS 1000) and polyvinyl pyrrolidone co-vinyl acetate 64 (PVPVA 64) or hydroxy propyl methyl cellulose 2910 (HPMC 2910) were developed in order to improve the dissolution of UC 781. UC 781 dissolution rate was markedly improved as compared to the physical mixtures and the pure drug, attaining maximum drug releases of up to 100% after only 5 min in the case of TPGS 1000-UC 781-PVPVA 64 solid dispersions and 30 min in TPGS 1000-UC 781-HPMC 2910. The increased UC 781 dissolution rate could be maintained when formulating UC 781 in PVPVA 64 tablets. The latter disintegrated in only 4 min, reaching drug releases of up to 90% (w/w). In addition, as opposed to the corresponding solid dispersions, no decrease in drug release occurred upon dissolution of PVPVA 64 tablets when the pH was increased to 6.8. Contrary to the PVPVA 64 tablet formulations, HPMC 2910 tablets showed a slow dissolution process due to the gelling nature of the polymer. The drug was slowly released as HPMC 2910 dissolved in the medium, however also in this case 90% (w/w) of the drug was dissolved after 4 h. Both polymers formed compatible blends in combination with the drug. Thermal analysis of the ternary mixtures revealed eutectic behavior exhibiting an extremely fine dispersion of the drug in the carrier. This was confirmed by the fact that no drug crystals could be detected using X-ray diffraction (XRD). As opposed to the physical mixtures, PVPVA 64 and HPMC 2910 solid dispersions did not contain any isolated polymer-rich phases, hence showed improved homogeneity. Amorphous TPGS 1000 clusters occurred in PVPVA 64 and HPMC 2910 formulations upon addition of at least 10% (w/w) UC 781, showing extremely low glass transition temperatures depending of the thermal history of the samples.


Assuntos
Anilidas/química , Fármacos Anti-HIV/química , Portadores de Fármacos , Furanos/química , Metilcelulose/análogos & derivados , Povidona/análogos & derivados , Vitamina E/análogos & derivados , Química Farmacêutica , Composição de Medicamentos , Derivados da Hipromelose , Cinética , Metilcelulose/química , Modelos Químicos , Polietilenoglicóis/química , Povidona/química , Pirrolidinas , Solubilidade , Comprimidos , Tecnologia Farmacêutica , Termodinâmica , Tioamidas , Temperatura de Transição , Compostos de Vinila , Vitamina E/química
8.
J Cyst Fibros ; 3 Suppl 2: 53-4, 2004 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-15463926

RESUMO

In this article, we describe a technique for culturing human airway epithelial cells, developed in Leuven, as a new tool for a most reliable diagnosis for ciliary disorders. This technique that allows to keep both structural and functional primary abnormalities of inherited ciliary abnormalities, while avoiding the secondary ones, can also be useful to a number of other studies, namely in cystic fibrosis.


Assuntos
Técnicas de Preparação Histocitológica/métodos , Mucosa Respiratória/patologia , Manejo de Espécimes/métodos , Técnicas de Cultura de Células , Células Epiteliais , Humanos , Coleta de Tecidos e Órgãos/métodos
9.
Eur J Pharm Biopharm ; 55(3): 283-9, 2003 May.
Artigo em Inglês | MEDLINE | ID: mdl-12754002

RESUMO

The aim of this study was to investigate the suitability of a sequential monolayer-suspension culture system as a model to screen subacute effects of drug excipients on ciliary beat frequency (CBF). The CBF of the cultured cells was measured by computerized microscope photometry. Protease inhibitors (puromycin, bestatin, bacitracin, actinonin and thiomersal) were used as model compounds and the mechanisms of ciliary inhibition were investigated by probing the involvement of arachidonic acid metabolism, guanylate cyclase (cGMP), protein kinase C (PKC) and adenosinetriphosphate (ATP) inhibition. Bestatin concentration-dependently reduced CBF by inhibiting arachidonic acid metabolism, cGMP, PKC and endogenous ATP consumption. Thiomersal and DMSO used for dissolving actinonin reduced CBF (P<0.05) via a non-specific mechanism. Bacitracin (8 mM) and puromycin (135 mM) had no effect on CBF after acute exposure (15-30 min) (P>0.05), but significantly reduced the CBF by approximately 15.0% following daily 15-min exposure for 1 week. This study shows that (i) sequential monolayer-suspension culture system is a valid model to screen both acute and subacute effects of drug excipients on CBF; and (ii) bacitracin, puromycin and actinonin are more cilio-compatible than bestatin and thiomersal and as such are more potentially useful nasal absorption enhancer from ciliotoxicity perspective.


Assuntos
Mucosa Nasal/citologia , Mucosa Nasal/efeitos dos fármacos , Inibidores de Proteases/farmacologia , Técnicas de Cultura de Células/métodos , Células Cultivadas , Cílios/efeitos dos fármacos , Cílios/enzimologia , Cílios/fisiologia , Humanos , Mecânica , Mucosa Nasal/enzimologia , Mucosa Nasal/fisiologia
10.
Int J Pharm ; 237(1-2): 179-91, 2002 Apr 26.
Artigo em Inglês | MEDLINE | ID: mdl-11955816

RESUMO

This study examined the potential usefulness of cultured human nasal epithelium as a model to investigate nasal absorption enhancement strategies for therapeutic peptides. The transport of leucine enkephalin (Leu-Enk) in the presence of bestatin and puromycin, respectively and various combinations of these protease inhibitors with absorption enhancers capable of inhibiting proteases or protecting peptides against protease degradation (glycocholate, dimethyl-beta-cyclodextrin (DM beta CD)) was studied. Epithelial membrane perturbation, protein leakage, bestatin/puromycin absorption and rebound aminopeptidase activity were used as toxicological end-points. The combination of puromycin with glycocholate or DM beta CD resulted in a higher absorption enhancement of Leu-Enk (9-14%) than when the absorption enhancers were combined with bestatin (1-3%) or when the inhibitors were used alone (2-4%). The higher absorption enhancement resulting from the combination of protease inhibitors with absorption enhancers caused a significant reduction of epithelial resistance and increased sodium fluorescein transport. Although only puromycin permeated the human nasal epithelium, both protease inhibitors induced a significant rebound aminopeptidase activity (25-61%), which can be associated with protein leakage (21-46%). This study highlighted (i) the potential usefulness of cultured human nasal epithelium as a model to study nasal absorption enhancement of therapeutic peptides; (ii) further studies using in vivo nasal models are required to ascertain whether the membrane perturbation and cytotoxicity observed with various combinations of the protease inhibitors and absorption enhancers really raise safety concerns.


Assuntos
Mucosa Nasal/metabolismo , Peptídeos/farmacocinética , Absorção/fisiologia , Administração Intranasal , Transporte Biológico , Células Cultivadas , Encefalina Leucina/metabolismo , Humanos , Mucosa Nasal/citologia , Peptídeos/uso terapêutico , Inibidores de Proteases/farmacocinética
11.
Acta Otolaryngol ; 124(4): 527-31, 2004 May.
Artigo em Inglês | MEDLINE | ID: mdl-15224888

RESUMO

OBJECTIVE: Ciliary orientation (COR) is an important parameter of mucociliary clearance and ciliary disorientation has been reported in cases of acquired abnormalities [secondary ciliary dyskinesia (SCD)] and in a very few cases as the single abnormality in primary ciliary dyskinesia (PCD). The etiology, pathogenesis, consequences and relevance of ciliary (dis)orientation are still unclear. MATERIAL AND METHODS: To elucidate the primary or secondary nature of ciliary (dis)orientation, COR was measured in 179 non-PCD and 59 PCD patients. COR was measured in biopsies and after ciliogenesis in culture and was correlated with a number of functional and ultrastructural parameters. COR was defined as the SD of the angles of lines through the central pair of microtubules using transmission electron microscopy. Internationally accepted normal values for COR are < or = 20 degrees; COR values of 20-35 degrees indicate increased disorientation; and COR values > 35 degrees represent a random orientation. RESULTS: For non-PCD biopsies, COR increased with increasing SCD, from 15 +/- 7 degrees (n = 54) for normal (< 5%) SCD to 28 +/- 8 degrees (n = 16) for severe (> 25%) SCD. No correlation was found between COR and ciliary beat frequency. However, increased COR values (28 +/- 8 degrees) were found for immotility (n = 8), compared to (coordinated) ciliary activity (19 +/- 9 degrees) (n = 121). After ciliogenesis no ultrastructural abnormalities were found and COR was normal (13 +/- 5 degrees; n = 308). COR can therefore be considered to be secondary in non-PCD and correlates with SCD percentage and ciliary motility. In biopsies from PCD patients with dynein deficiency and with normal ultrastructure, COR was increased, to 28 +/- 11 degrees (n = 32) and 21 +/- 7 degrees (n = 15), respectively, and in cases with central pair abnormalities COR was random (38 +/- 11 degrees; n = 12). After ciliogenesis COR remained random in the PCD group with central pair abnormalities (38 +/- 9 degrees; n = 15), and was increased in the PCD groups with dynein deficiency (24 +/- 10 degrees; n = 35) and normal ultrastructure (25 +/- 8 degrees; n = 17). Ciliary disorientation was never found as the single abnormality. CONCLUSION: COR can be considered to be secondary in PCD. Both ciliary (im)motility and SCD percentage contribute to COR.


Assuntos
Cílios/ultraestrutura , Transtornos da Motilidade Ciliar/patologia , Mucosa Respiratória/patologia , Cílios/fisiologia , Transtornos da Motilidade Ciliar/fisiopatologia , Técnicas de Cultura , Humanos , Síndrome de Kartagener/patologia , Síndrome de Kartagener/fisiopatologia , Depuração Mucociliar
12.
Vaccine ; 30(40): 5849-55, 2012 Aug 31.
Artigo em Inglês | MEDLINE | ID: mdl-22824343

RESUMO

BACKGROUND: Foot-and-mouth disease (FMD) vaccine potency testing involves hundreds of animals each year. Despite considerable efforts during the past decades, a challenge-free alternative vaccine potency test to replace the European protective dose 50% test (PD(50)) has not been implemented yet. The aim of the present study was to further characterize the properties of serological vaccine potency models. METHODS: Logistic regression models were built for 5 serological assays from 3 different laboratories. The serum samples originated from 5 repeated PD(50) vaccine potency trials with a highly potent A/IRN/11/96 vaccine. Receiver Operating Characteristic analysis was used to determine a serological pass mark for predicting in vivo protected animals. Subsequently, an estimated PD(50) was calculated and the serotype dependency of the logistic models was investigated. RESULTS: Although differences were observed between the laboratories and the serological assays used, the logistic models accurately predicted the in vivo protection status of the animals in 74-93% of the cases and the antibody pass levels corresponded to 84-97% of protection, depending on the serological assay used. For logistic models that combine different serotypes, the model fit can be increased by inclusion of a serotype factor in the logistic regression function. CONCLUSIONS: The in vitro estimated PD(50) method may be at least as precise as the in vivo PD(50) test and may accurately predict the PD(50) content of a vaccine. However, the laboratory-effect and the serotype-dependency should be further investigated.


Assuntos
Febre Aftosa/prevenção & controle , Modelos Logísticos , Vacinas Virais/imunologia , Animais , Anticorpos Antivirais/sangue , Bovinos , Ensaio de Imunoadsorção Enzimática , Vírus da Febre Aftosa , Masculino , Testes de Neutralização , Valor Preditivo dos Testes , Curva ROC , Reprodutibilidade dos Testes , Estatística como Assunto , Vacinas Virais/normas
13.
J Virol Methods ; 173(1): 108-14, 2011 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-21295609

RESUMO

Cell-based assays are still used widely in foot-and-mouth disease (FMD) research, despite the existence of a wide variety of molecular techniques. The aim of this study was to validate an automated, quantitative spectrometric reading to replace the time-consuming and subjective microscopic (MIC) evaluation of the FMD virus-induced cytopathic effect (CPE). Therefore, the diagnostic performance of two commercial cell viability assays (CellTiter 96(®) AQueous One Solution Cell Proliferation Assay (MTS) and CellTiter-Blue(®) Cell Viability Assay (CTB), both from Promega, Leiden, The Netherlands) was evaluated. Following optimization of the assay protocols and using the MIC results as a reference standard, the absorbance-read MTS assay, the fluorescence-read CTB assay and the absorbance-read CTB (CTB(abs)) assay demonstrated similar high sensitivities (97%, 99% and 98%, respectively), specificities (100%, 98% and 99%, respectively), accuracy measures (0.99, 0.98 and 0.98, respectively), precision measures (1.00, 0.98 and 0.99, respectively) and Cohen kappa agreement indices (0.97, 0.97 and 0.96, respectively) for detecting CPE in cell cultures. Due to its performance, cost effectiveness and ease of use, the CTB(abs) assay was selected for further evaluation of its ability to detect virus neutralization and to screen antiviral compounds. The CTB(abs) assay had 99% sensitivity and 100% specificity for the detection of neutralizing antibodies in sera from cattle infected with FMDV and in sera from unvaccinated, uninfected cattle and resulted in a mean Z'-factor of 0.85 for antiviral compound test plates. The CTB(abs) assay is now used routinely in the Belgian FMD reference laboratory for serological testing and high-throughput antiviral compound screening.


Assuntos
Anticorpos Neutralizantes/sangue , Anticorpos Antivirais/sangue , Doenças dos Bovinos/imunologia , Vírus da Febre Aftosa/imunologia , Febre Aftosa/imunologia , Virologia/métodos , Animais , Automação/métodos , Bovinos , Sobrevivência Celular , Colorimetria/métodos , Fluorometria/métodos , Sensibilidade e Especificidade
14.
Vaccine ; 27(5): 741-7, 2009 Jan 29.
Artigo em Inglês | MEDLINE | ID: mdl-19041355

RESUMO

The selection of matching strains for use in outbreaks of foot-and-mouth disease (FMD) virus can be assessed in vivo or by serological r-value determination. Sera from animals involved in vaccine potency and cross-protection trials performed using the "Protection against Podal Generalization" (PPG) test for two serotype A strains were collected and analyzed by the virus neutralization test (VNT) and liquid-phase ELISA (lpELISA) in three laboratories. The average VNT r-values for medium and high serum titer classes from the A(24) Cruzeiro vaccinated animals were in line with the A/Arg/01 heterologous PPG outcome for all testing laboratories, suggesting that the vaccine strain A(24) Cruzeiro is unlikely to protect against the field isolate A/Arg/01. The corresponding lpELISA r-values were slightly higher and indicate a closer relationship between both strains. Pooling of serum samples significantly reduced the inter-animal and inter-trial variation. The results suggest that a suitable reference serum for vaccine matching r-value experiments might be a pool or a medium to high VNT or lpELISA titer serum. Furthermore, the VNT seems to produce the most reproducible inter-laboratory results. More work is, however, needed in order to substantiate these claims.


Assuntos
Vírus da Febre Aftosa/classificação , Vírus da Febre Aftosa/imunologia , Febre Aftosa/prevenção & controle , Febre Aftosa/virologia , Vacinas Virais/imunologia , Animais , Anticorpos Antivirais/sangue , Anticorpos Antivirais/imunologia , Argentina/epidemiologia , Bovinos , Reações Cruzadas , Surtos de Doenças , Ensaio de Imunoadsorção Enzimática , Febre Aftosa/epidemiologia , Imunoensaio/normas , Testes de Neutralização
15.
Vaccine ; 26(31): 3870-9, 2008 Jul 23.
Artigo em Inglês | MEDLINE | ID: mdl-18555565

RESUMO

Foot-and-mouth disease (FMD) vaccine potency testing has historically been performed by experimentally infecting vaccinated cattle. A few alternative approaches to the in vivo challenge test based on the correlation between serum titres of primo-vaccinated cattle and protection against infection have been proposed, but none have been accepted by the European Pharmacopoeia (Ph.Eur.) due to the lack of statistical power and the pooling of data over time. The present study addresses these issues and presents data of 150 cattle vaccinated according to Ph.Eur. standards. Four laboratories took part in the serological testing and different serological assays were used, including virus neutralisation assays and ELISA formats. Models correlating specific anti-FMD virus antibody titres to protection were built using logistic regression followed by Receiver Operating Characteristic (ROC) analysis. The best models accurately predicted the in vivo protection status in 80.0% of the cases. Although differences were observed between laboratories and assays used, the majority of antibody pass-levels, determined using ROC analysis, corresponded to at least 75.0% probability of protection. The indirect potency assessment procedure proposed is at least as precise (repeatability=65.8%, reproducibility=60.7%) as the in vivo test, can be standardised and results in a quantitative PD50 value. The validity of the procedure was also demonstrated.


Assuntos
Anticorpos Antivirais/sangue , Febre Aftosa/prevenção & controle , Vacinas Virais/imunologia , Animais , Bovinos , Ensaio de Imunoadsorção Enzimática , Modelos Logísticos , Testes de Neutralização , Valor Preditivo dos Testes , Curva ROC , Reprodutibilidade dos Testes , Estatística como Assunto
16.
Pharm Res ; 20(8): 1125-32, 2003 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-12948008

RESUMO

PURPOSE: The purpose of this study was to provide functional and molecular evidence to support the existence of large neutral amino acid transporters in human nasal epithelium using nasal primary cell culture model. METHODS: L-Phenylalanine was used as a model substrate to characterize carrier-mediated permeation of amino acids across human nasal epithelium. The influence of temperature, concentration, other amino acids, metabolic/transport inhibitors, and polarity/stereo-selectivity on transport of the model compound was investigated. Reverse transcriptase polymerase chain reaction was used for molecular characterization of the existence of the transporters. RESULTS: The transport of L-phenylalanine across the human nasal epithelium was polarized (apical --> basolateral >> basolateral --> apical), saturable (Km = 1.23 mM; Vmax = 805.1 nmol/mg protein/min) and stereo-selective (permeation of L-phenylalanine >> D-Phenylalanine). Its permeation was significantly (< 0.05) reduced by cationic, small and large neutral amino acids, oubain, amiloride, sodium-free medium, and temperature lowering. Reverse transcriptase polymerase chain reaction revealed the presence of the broad-scope cationic-dependent amino acid transporter gene (y+LAT-2) in the human nasal epithelium. CONCLUSIONS: Based on the results of this study, one may postulate that the human nasal epithelium expresses L-amino acid transporters. More studies are necessary for detailed characterization of the transporters.


Assuntos
Sistemas de Transporte de Aminoácidos/metabolismo , Mucosa Nasal/metabolismo , Fenilalanina/metabolismo , Sistemas de Transporte de Aminoácidos/genética , Animais , Transporte Biológico Ativo , Células Cultivadas , Humanos , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Temperatura
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