RESUMO
This study was set up to assess the performance of the Reveal® rapid AST system to determine the drug susceptibility of Pseudomonas aeruginosa strains directly from blood cultures. Two hundred fully sequenced clinical P. aeruginosa strains were selected for the evaluation, of which 26.5% (n = 53) produced transferable ß-lactamases, and 2.0 to 33.0% had susceptibility levels close to the EUCAST 2021 breakpoints of 11 commonly used antipseudomonal antibiotics. The Reveal® AST system was run with a commercial MIC microplate designed for fast-growing Gram-negative bacilli (Microscan Neg MDR MIC 1), and was compared to the manually operated GN6F MIC microdilution panel from Thermo Fisher, as a comparator method. The Reveal® AST system provided MIC results for the 11 antipseudomonal antibiotics tested within a mean time to result of 6 h 22 min. By comparison with the GN6F panel, the overall rates of categorical agreement (CA), very major errors (VME), major errors (ME), and minor errors (mE for meropenem only) were 96.1%, 1.6%, 4.2%, and 0.6%, respectively. The Specific Reveal® AST system appears to be a reliable and fast technology to determine the susceptibility of P. aeruginosa to antibiotics, including those with resistance levels near categorical breakpoints, directly from blood cultures.
Assuntos
Hemocultura , Pseudomonas aeruginosa , Humanos , Hemocultura/métodos , Testes de Sensibilidade Microbiana , Antibacterianos/farmacologia , Bactérias Gram-NegativasRESUMO
OBJECTIVE: To determine a personalized and optimized contrast injection protocol for a uniform and optimal diagnostic level of liver parenchymal enhancement, in a large patient population enrolled in a multicenter study. METHODS: Six hundred ninety-two patients who underwent a standardized multi-phase liver CT examination were prospectively assigned to one contrast media (CM) protocol group: G1 (100 mL fixed volume, 37 gI); G2 (600 mgI/kg of total body weight (TBW)); G3 (750 mgI/kg of fat-free mass (FFM)), and G4 (600 mgI/kg of FFM). Change in liver parenchyma CT number between unenhanced and contrast-enhanced images was measured by two radiologists, on 3-mm pre-contrast and portal phase axial reconstructions. The enhancement histograms were compared across CM protocols, specifically according to a target diagnostic value of 50 HU. The total amount of iodine dose was also compared among protocols by median and interquartile range (IQR). The Kruskal-Wallis and Mann-Whitney U tests were used to assess significant differences (p < 0.005), as appropriate. RESULTS: A significant difference (p < 0.001) was found across the groups with liver enhancement decreasing from median over-enhanced values of 77.0 (G1), 71.3 (G2), and 65.1 (G3) to a target enhancement of 53.2 HU for G4. Enhancement IQR was progressively reduced from 26.5 HU (G1), 26.0 HU (G2), and 17.8 HU (G3) to 14.5 HU (G4). G4 showed a median iodine dose of 26.0 gI, significantly lower (p < 0.001) than G3 (33.9 gI), G2 (38.8 gI), and G1 (37 gI). CONCLUSIONS: The 600 mgI/kg FFM-based protocol enabled a diagnostically optimized liver enhancement and improved patient-to-patient enhancement uniformity, while significantly reducing iodine load. KEY POINTS: ⢠Consistent and clinically adequate liver enhancement is observed with personalized and optimized contrast injection protocol. ⢠Fat-free mass is an appropriate body size parameter for correlation with liver parenchymal enhancement. ⢠Diagnostic oncology follow-up liver CT examinations may be obtained using 600 mgI/kg of FFM.
Assuntos
Meios de Contraste , Iodo , Humanos , Fígado/diagnóstico por imagem , Estudos Multicêntricos como Assunto , Estudos Prospectivos , Tomografia Computadorizada por Raios XRESUMO
Mutation-dependent overproduction of intrinsic ß-lactamase AmpC is considered the main cause of resistance of clinical strains of Pseudomonas aeruginosa to antipseudomonal penicillins and cephalosporins. Analysis of 31 AmpC-overproducing clinical isolates exhibiting a greater resistance to ceftazidime than to piperacillin-tazobactam revealed the presence of 17 mutations in the ß-lactamase, combined with various polymorphic amino acid substitutions. When overexpressed in AmpC-deficient P. aeruginosa 4098, the genes coding for 20/23 of these AmpC variants were found to confer a higher (2-fold to >64-fold) resistance to ceftazidime and ceftolozane-tazobactam than did the gene from reference strain PAO1. The mutations had variable effects on the MICs of ticarcillin, piperacillin-tazobactam, aztreonam, and cefepime. Depending on their location in the AmpC structure and their impact on ß-lactam MICs, they could be assigned to 4 distinct groups. Most of the mutations affecting the omega loop, the R2 domain, and the C-terminal end of the protein were shared with extended-spectrum AmpCs (ESACs) from other Gram-negative species. Interestingly, two new mutations (F121L and P154L) were predicted to enlarge the substrate binding pocket by disrupting the stacking between residues F121 and P154. We also found that the reported ESACs emerged locally in a variety of clones, some of which are epidemic and did not require hypermutability. Taken together, our results show that P. aeruginosa is able to adapt to efficacious ß-lactams, including the newer cephalosporin ceftolozane, through a variety of mutations affecting its intrinsic ß-lactamase, AmpC. Data suggest that the rates of ESAC-producing mutants are ≥1.5% in the clinical setting.
Assuntos
Adaptação Fisiológica/genética , Antibacterianos/farmacologia , Proteínas de Bactérias/genética , Mutação , Pseudomonas aeruginosa/genética , Resistência beta-Lactâmica/genética , beta-Lactamases/genética , Adaptação Fisiológica/efeitos dos fármacos , Sequência de Aminoácidos , Substituição de Aminoácidos , Aztreonam/farmacologia , Proteínas de Bactérias/metabolismo , Cefepima , Ceftazidima/farmacologia , Cefalosporinas/farmacologia , Expressão Gênica , Testes de Sensibilidade Microbiana , Dados de Sequência Molecular , Ácido Penicilânico/análogos & derivados , Ácido Penicilânico/farmacologia , Piperacilina/farmacologia , Combinação Piperacilina e Tazobactam , Estrutura Secundária de Proteína , Estrutura Terciária de Proteína , Pseudomonas aeruginosa/efeitos dos fármacos , Pseudomonas aeruginosa/metabolismo , Tazobactam , Ticarcilina/farmacologia , beta-Lactamases/metabolismoRESUMO
We observe apparent hole pockets in the Fermi surfaces of single-layer Bi-based cuprate superconductors from angle-resolved photoemission. From detailed low-energy electron diffraction measurements and an analysis of the angle-resolved photoemission polarization dependence, we show that these pockets are not intrinsic but arise from multiple overlapping superstructure replicas of the main and shadow bands. We further demonstrate that the hole pockets reported recently from angle-resolved photoemission [Meng et al., Nature (London) 462, 335 (2009)] have a similar structural origin and are inconsistent with an intrinsic hole pocket associated with the electronic structure of a doped CuO2 plane.
RESUMO
OBJECTIVES: To compare deep learning (True Fidelity, TF) and partial model based Iterative Reconstruction (ASiR-V) algorithm for image texture, low contrast lesion detectability and potential dose reduction. METHODS: Anthropomorphic phantoms (mimicking non-overweight and overweight patient), containing lesions of 6â¯mm in diameter with 20HU contrast, were scanned at five different dose levels (2,6,10,15,20â¯mGy) on a CT system, using clinical routine protocols for liver lesion detection. Images were reconstructed using ASiR-V 0% (surrogate for FBP), 60 % and TF at low, medium and high strength. Noise texture was characterized by computing a normalized Noise Power Spectrum filtered by an eye filter. The similarity against FBP texture was evaluated using peak frequency difference (PFD) and root mean square deviation (RMSD). Low contrast detectability was assessed using a channelized Hotelling observer and the area under the ROC curve (AUC) was used as figure of merit. Potential dose reduction was calculated to obtain the same AUC for TF and ASiR-V. RESULTS: FBP-like noise texture was more preserved with TF (PFD from -0.043mm-1 to -0.09mm-1, RMSD from 0.12mm-1 to 0.21mm-1) than with ASiR-V (PFD equal to 0.12 mm-1, RMSD equal to 0.53mm-1), resulting in a sharper image. AUC was always higher with TF than ASIR-V. In average, TF compared to ASiR-V, enabled a radiation dose reduction potential of 7%, 25 % and 33 % for low, medium and high strength respectively. CONCLUSION: Compared to ASIR-V, TF at high strength does not impact noise texture and maintains low contrast liver lesions detectability at significant lower dose.
Assuntos
Aprendizado Profundo , Neoplasias Hepáticas , Algoritmos , Humanos , Neoplasias Hepáticas/diagnóstico por imagem , Imagens de Fantasmas , Doses de Radiação , Interpretação de Imagem Radiográfica Assistida por Computador , Tomografia Computadorizada por Raios XRESUMO
We report a method based on heterodyne numerical holography associated to photothermal excitation for full field and three-dimensional localisation of metallic nanoparticles. A modulated pump laser (lambda = 532 nm) heats several particles, creating local refractive index changes. This modulation is detected using a probe and a local oscillator beam (lambda = 785 nm), frequency-shifted to create a hologram beating at low frequency. Tens of particles, down to diameters of 10 nm, can be localised simultaneously and selectively in three dimensions with near- diffraction resolution by a numerical reconstruction of a single hologram acquired in 5 s.
Assuntos
Ouro/química , Holografia/métodos , Nanopartículas/química , Nanopartículas/ultraestrutura , Fotometria/métodos , Refratometria/métodos , Termografia/métodosRESUMO
Patterns of summertime movement and habitat use of yellow-stage American eels Anguilla rostrata within York River and estuary and Gaspé Bay (Gaspesia, Québec, Canada) were examined using acoustic telemetry. Fifty fish were tagged with acoustic transmitters and released, either in the river or in the upper estuary, and their patterns of movement and habitat use were monitored at short spatial and temporal scales during the summer months using a dense hydrophone array. Approximately half of the fish released in the river swam to the estuary; two-thirds of the fish released within the estuary did not move out of the estuary. Anguilla rostrata were detected more frequently and had a greater areal range of detections during night, suggesting greater nocturnal activity. Longitudinal movements within the estuary tended to occur nocturnally, with upstream movements from early to late evening, and downstream movements from late evening to early morning. Approximately one-third of fish showed a regular pattern of movement, tending to reside in the deeper, downstream part of the estuary during day and in the shallower, more upstream part of the estuary during night. Approximately a quarter of fish, located in the upper estuary, remained upstream during both night and day. The remaining fish showed patterns intermediate between these two.
Assuntos
Acústica/instrumentação , Anguilla/fisiologia , Telemetria/métodos , Animais , Fotoperíodo , Quebeque , Rios , Estações do AnoRESUMO
INTRODUCTION: Little is known about the functional symptoms associated with Lyme borreliosis (LB) in Europe. We aimed to assess functional symptoms associated with presumed LB and to compare patients with and without confirmed LB. MATERIALS AND METHODS: We performed a retrospective monocenter study. Patients consulting for presumed LB were included. RESULTS: Between November 2015 and June 2018, 355patients were included (mean age: 51years, 52% of women) of which 48had LB: erythema migrans (42%), early disseminated LB (50%; 35% of neuroborreliosis cases), and late disseminated LB (8%). The most frequently reported functional symptoms were neuropathic pain (23%), arthralgia (23%), and asthenia (17%). Other functional symptoms were rare (≤10%). Three hundred and seven (86%) patients did not have LB. Patients with confirmed LB reported fewer functional symptoms than patients without LB (1.8 (±1.7) vs. 3.6 (±2.5), P<0.001) with a shorter duration of symptoms (< 3 months in 48% vs. 16% of cases, P<0.001). They less often reported asthenia (17% vs. 59%, P<0.001), widespread pain (10% vs. 31%, P=0.003), myalgia (10% vs. 32%, P=0.002), memory disorders (4% vs. 16%, P=0.03), irritability (2% vs. 23%, P±0.001), and sadness (0% vs. 16%, P=0.003). CONCLUSION: In patients consulting for presumed LB, patients diagnosed with LB had fewer and shorter functional symptoms than patients without LB.
Assuntos
Doença de Lyme/diagnóstico , Avaliação de Sintomas , Adulto , Idoso , Antibacterianos/uso terapêutico , Borrelia burgdorferi/imunologia , Borrelia burgdorferi/isolamento & purificação , Diagnóstico Diferencial , Eritema Migrans Crônico/diagnóstico , Eritema Migrans Crônico/epidemiologia , Eritema Migrans Crônico/terapia , Feminino , França/epidemiologia , Humanos , Doença de Lyme/epidemiologia , Doença de Lyme/terapia , Neuroborreliose de Lyme/diagnóstico , Neuroborreliose de Lyme/epidemiologia , Neuroborreliose de Lyme/terapia , Masculino , Pessoa de Meia-Idade , Encaminhamento e Consulta , Estudos Retrospectivos , Testes Sorológicos , Resultado do TratamentoRESUMO
Breast-conserving surgery followed by radiotherapy is effective in reducing recurrence; however, telangiectasia and fibrosis can occur as late skin side effects. As radiotherapy acts through producing DNA damage, we investigated whether genetic variation in DNA repair and damage response confers increased susceptibility to develop late normal skin complications. Breast cancer patients who received radiotherapy after breast-conserving surgery were examined for late complications of radiotherapy after a median follow-up time of 51 months. Polymorphisms in genes involved in DNA repair (APEX1, XRCC1, XRCC2, XRCC3, XPD) and damage response (TP53, P21) were determined. Associations between telangiectasia and genotypes were assessed among 409 patients, using multivariate logistic regression. A total of 131 patients presented with telangiectasia and 28 patients with fibrosis. Patients with variant TP53 genotypes either for the Arg72Pro or the PIN3 polymorphism were at increased risk of telangiectasia. The odds ratios (OR) were 1.66 (95% confidence interval (CI): 1.02-2.72) for 72Pro carriers and 1.95 (95% CI: 1.13-3.35) for PIN3 A2 allele carriers compared with non-carriers. The TP53 haplotype containing both variant alleles was associated with almost a two-fold increase in risk (OR 1.97, 95% CI: 1.11-3.52) for telangiectasia. Variants in the TP53 gene may therefore modify the risk of late skin toxicity after radiotherapy.
Assuntos
Neoplasias da Mama/genética , Neoplasias da Mama/radioterapia , Dano ao DNA/genética , Reparo do DNA/genética , Polimorfismo Genético , Lesões por Radiação/genética , Adulto , Idoso , Idoso de 80 Anos ou mais , Neoplasias da Mama/cirurgia , Terapia Combinada/efeitos adversos , Dano ao DNA/fisiologia , Feminino , Seguimentos , Genes p53 , Haplótipos , Humanos , Desequilíbrio de Ligação , Mastectomia Segmentar/reabilitação , Pessoa de Meia-Idade , Polimorfismo Genético/fisiologia , Polimorfismo de Nucleotídeo Único , Lesões por Radiação/complicações , Lesões por Radiação/patologia , Dermatopatias/etiologia , Dermatopatias/genéticaRESUMO
Our hypothesis was that immediate repetition of a microsurgery-suturing task will improve its execution and outcome. This was an experimental animal study. Ten surgeons were divided into two groups of five surgeons. Each performed two end-to-end carotid anastomoses on the same rat, one after the other. The anastomosis was evaluated by the surgeon and an instructor. The primary endpoint was permeability. The outcome was evaluated using an objective and subjective assessment grid yielding 1 to 3 points per item. The total scores for each of the 10 surgeons were used to compare the anastomosis of carotid 1 versus 2, using the ratings given by the surgeon and the instructor. Twenty anastomoses were performed, but 1 rat died intraoperatively, leaving 18 anastomoses for evaluation. No significant differences were found on the main endpoint of permeability, with all anastomoses being permeable. The surgeon's self-assessment was significantly better for the second carotid artery (P=0.05), but this was not confirmed by the proxy assessment (instructor). The analysis by subgroups-morning versus afternoon-found the second carotid anastomosis was significant better in the self-assessment and proxy assessment for the morning group (P<0.001, P=0.024). There was no significant difference in clamping times. The immediate repetition of a microsurgical procedure seems to favor its execution, which leads us to propose that the more difficult or important anastomosis should be done after an easier or less important one during complex surgeries. LEVEL OF EVIDENCE: 2B.
Assuntos
Anastomose Cirúrgica/educação , Competência Clínica , Educação Médica Continuada/métodos , Microcirurgia/educação , Cirurgiões , Suturas , Animais , Artérias Carótidas/cirurgia , Humanos , Ratos Sprague-Dawley , Grau de Desobstrução VascularRESUMO
Cr2N is commonly found as a minority phase or inclusion in stainless steel, CrN-based hard coatings, etc. However, studies on phase-pure material for characterization of fundamental properties are limited. Here, Cr2N thin films were deposited by reactive magnetron sputtering onto (0001) sapphire substrates. X-ray diffraction and pole figure texture analysis show Cr2N (0001) epitaxial growth. Scanning electron microscopy imaging shows a smooth surface, while transmission electron microscopy and X-ray reflectivity show a uniform and dense film with a density of 6.6 g cm-3, which is comparable to theoretical bulk values. Annealing the films in air at 400 °C for 96 h shows little signs of oxidation. Nano-indentation shows an elastic-plastic behavior with H = 18.9 GPa and E r = 265 GPa. The moderate thermal conductivity is 12 W m-1 K-1, and the electrical resistivity is 70 µΩ cm. This combination of properties means that Cr2N may be of interest in applications such as protective coatings, diffusion barriers, capping layers and contact materials.
RESUMO
OBJECTIVES: Time to blood culture positivity (TTP), a routinely available parameter in automated blood culture systems, may be a proxy for infectious burden in patients with bloodstream infections. We aimed to study the association between TTP and infective endocarditis (IE), or death, in patients with Staphylococcus aureus bacteraemia. METHODS: VIRSTA is a multicentre prospective cohort study that included all adult patients with S. aureus bacteraemia in eight university hospitals in France (2009-2011). We analysed data from four centres which collected data on TTP. Regression models were used to study the association between TTP and definite IE (Duke-Li criteria), and 30 day-mortality. RESULTS: We included 587 patients with S. aureus bacteraemia: mean age was 65.3 ± 16.3 years, 420 out of 587 patients (71.6%) were male, 121 out of 587 (20.6%) died, and 42 out of 587 (7.2%) had definite IE. Median TTP of first positive blood culture was 13.7 h (interquartile range 9.9-18). On multivariate analysis, 30-day mortality was associated with TTP ≤13.7 h (74/295 (25.1%) vs. 47/292 (16.1%), p 0.02), as well as old age, McCabe score, methicillin resistance, stroke, pneumonia, and C-reactive protein. TTP was also independently associated with IE, but with a U-shape curve: IE was more common in the first (TTP <10 h, 17/148, 11.5%), and the last (TTP ≥18 h, 8/146, 5.5%) quartiles of TTP, p 0.002. CONCLUSIONS: TTP provides reliable information in patients with S. aureus bacteraemia, on the risk of IE, and prognosis, with short TTP being an independent predictor of death. These data, readily available at no cost, may be used to identify patients who require specific attention.
Assuntos
Bacteriemia/sangue , Bacteriemia/diagnóstico , Hemocultura/estatística & dados numéricos , Endocardite Bacteriana/mortalidade , Staphylococcus aureus Resistente à Meticilina/isolamento & purificação , Infecções Estafilocócicas/sangue , Infecções Estafilocócicas/diagnóstico , Idoso , Bacteriemia/microbiologia , Endocardite Bacteriana/diagnóstico , Endocardite Bacteriana/microbiologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Infecções Estafilocócicas/microbiologia , Fatores de TempoRESUMO
In social Hymenoptera, within-colony relatedness is usually high due to the haplodiploid sex-determining system. However, factors such as the presence of multiple reproductive queens (polygyny), multiple queen matings (polyandry) or worker reproduction result in decreased relatedness among workers and the brood they rear, and consequently dilute their inclusive fitness benefits from helping. Here, we investigated population genetic structure, mating system, worker reproduction and parthenogenesis in the desert ant Cataglyphis sabulosa. Analysis of worker genotypes showed that colonies are headed by a single queen, mated with 1-5 males. The inbreeding coefficient within colonies and the levels of relatedness between the queens and their mates were positive, indicating that mating occurs between related individuals. Moreover, the mates of a queen are on average related and contribute equally to worker production. Our analyses also indicate that colonies are genetically differentiated and form a population exhibiting no isolation-by-distance pattern, consistent with the independent foundation of new colonies (that is, without the help of workers). Finally, both ovarian dissections and genetic data on the parentage of males show that workers do not reproduce in queenright colonies; however, they lay both haploid (arrhenotokous males) and diploid (thelytokous females) eggs in queenless colonies. In contrast to the congeneric species C. cursor, where new queens are produced by thelytokous parthenogenesis, female sexuals of C. sabulosa result from classical sexual reproduction.
Assuntos
Formigas/genética , Animais , Formigas/fisiologia , Feminino , Masculino , PartenogêneseRESUMO
Pre-steady-state catalytic properties of insect acetylcholinesterase (AChE, EC 3.1.1.7) were studied with the neutral substrate N-methylindoxylacetate. Kinetics of soluble Apis mellifera and Drosophila melanogaster AChE forms showed lags (v(i)=0) before reaching the steady-state. Results were interpreted in terms of slow equilibrium between two conformational states E and E' of insect AChE. Hysteresis of insect AChE has been pointed out for the first time. The hysteretic behaviour was found to depend on the NMIA concentration and the nature of the enzyme. The maximum induction times (tau(max)) to reach the steady-state were 800 and 1000s with soluble AChE from A. mellifera and D.melanogaster, respectively. The orders of magnitude of the tau(max) were high and similar to human AChE and BuChE.
Assuntos
Acetilcolinesterase/metabolismo , Abelhas/enzimologia , Drosophila melanogaster/enzimologia , Animais , Biocatálise , Células CHO , Cricetinae , Cricetulus , Humanos , Hidrólise , Cinética , Proteínas Recombinantes/metabolismoRESUMO
Type 1 17beta-hydroxysteroid dehydrogenase (17beta-HSD1) is a key steroidogenic enzyme that catalyses the reduction of steroid estrone into the most potent endogenous estrogen estradiol using the cofactor NAD(P)H. Bisubstrate inhibition is a good way to enhance the potency of inhibitors of cofactor-assisted enzymes. The design of a bisubstrate inhibitor of 17beta-HSD1, the estradiol/adenosine hybrid EM-1745, is reviewed and strategies for future designs of inhibitors are proposed.
Assuntos
17-Hidroxiesteroide Desidrogenases/antagonistas & inibidores , Desenho de Fármacos , Inibidores Enzimáticos/química , Inibidores Enzimáticos/farmacologia , 17-Hidroxiesteroide Desidrogenases/química , 17-Hidroxiesteroide Desidrogenases/metabolismo , Cristalização , Humanos , Especificidade por SubstratoRESUMO
The authors report a case of septic pericardial effusion resulting in cardiac tamponade associated with intrathoracic botryomycosis in a dog. Septic pericarditis and a pulmonary mass were diagnosed, and subtotal pericardiectomy and lobectomy of the affected pulmonary areas were carried out. Histopathology of the excised tissue showed changes supportive of botryomycosis--namely a pyogranulomatous inflammation with neutrophils centred around amorphous homogeneous eosinophilic material and club-like bodies containing Gram-positive bacterial cocci present in the centre. The patient recovered well following surgery and antibiotic therapy. To the authors' knowledge, this is the first report of pulmonary botryomycosis in the dog and the first report of this condition presented with pericardial involvement and cardiac tamponade in any species.
Assuntos
Doenças do Cão/diagnóstico , Pneumopatias Fúngicas/veterinária , Derrame Pericárdico/veterinária , Pericardite/veterinária , Infecções Estafilocócicas/veterinária , Animais , Tamponamento Cardíaco/diagnóstico , Tamponamento Cardíaco/cirurgia , Tamponamento Cardíaco/veterinária , Doenças do Cão/cirurgia , Cães , Pneumopatias Fúngicas/diagnóstico , Pneumopatias Fúngicas/cirurgia , Masculino , Derrame Pericárdico/diagnóstico , Derrame Pericárdico/cirurgia , Pericardiectomia/métodos , Pericardiectomia/veterinária , Pericardite/diagnóstico , Pericardite/cirurgia , Infecções Estafilocócicas/diagnóstico , Infecções Estafilocócicas/cirurgia , Resultado do TratamentoRESUMO
A new MALDI-TOF MS imipenem hydrolysis assay was developed to optimize detection of carbapenemase-producing strains of Pseudomonas aeruginosa. The method correctly discriminated carbapenemase producers (n=74) from non-producers (n=95) in 99.4% of cases, and successfully detected 100% GES-5 strains (n=5).
Assuntos
Proteínas de Bactérias/metabolismo , Infecções por Pseudomonas/diagnóstico , Pseudomonas aeruginosa/enzimologia , Pseudomonas aeruginosa/isolamento & purificação , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos , beta-Lactamases/metabolismo , Membrana Celular/efeitos dos fármacos , Permeabilidade da Membrana Celular/efeitos dos fármacos , Colistina/farmacologia , Humanos , Imipenem/metabolismo , Infecções por Pseudomonas/microbiologia , Pseudomonas aeruginosa/efeitos dos fármacos , Sensibilidade e EspecificidadeRESUMO
We demonstrate here a simple measurement protocol which allows the thermal properties of anisotropic crystalline materials to be determined. This protocol is validated by the measurement of Bi2Se3, a layered material consisting of covalently bonded sheets with weak van der Waals bonds between each layer, which has highly anisotropic thermal properties. Thermoreflectance microscopy measurements were carried out on a single-crystal Bi2Se3 sample, firstly on the bare sample and then after capping with a 100 nm thick gold layer. Whereas on the bare sample lateral heat diffusion is dominated by the in-plane thermal diffusivity, on the metal-capped substrate heat diffusion perpendicular to the sample surface dominates. Using a simple theoretical model, we show how this double measurement protocol allows the anisotropic thermal conductivity coefficients of bulk Bi2Se3 to be evaluated.
RESUMO
Structural relationships between colonic mucin species were assessed using a library of monoclonal antibodies (MAbs) directed against purified human colonic mucin (HCM). After immunization of mice with purified mucin from normal human colonic mucosa, 14% of 1,920 fusion products screened were positive for anti-HCM activity in a solid-phase assay. Patterns of selective binding by hybridomas to six discrete HCM species (I-VI) separated by DEAE-cellulose chromatography suggested the presence of both shared and species-specific antigenic determinants among HCM species I-VI. 23 anti-HCMs MAbs (7 IgM, 7 IgG1, and 9 IgG2) demonstrating a range of anti-HCM species specificities, were produced and used to study structural relationships between mucin species. Binding of various mucin species by individual anti-HCM MAbs was shown by competitive solid-phase radioimmunoassay to reflect the presence of identical epitopes on the different species. Adsorption of HCM species on a variety of affinity resins prepared with anti-HCM MAbs demonstrated that binding to multiple mucin species by a single MAb was related to intrinsic structural determinants. Four anti-HCM MAbs recognized protease-sensitive antigenic structures, which suggests that they may be directed to core HCM proteins. 12 of the anti-HCM MAbs were shown by solid-phase assay to recognize either complete (n = 5) or partial (n = 7) isolated colonic mucin oligosaccharide side chains of defined structure. Collectively, these data show the presence of both shared and unique antigenic structural determinants among colonic mucin species. Chromatographic heterogeneity of mucin glycoproteins seems to be related to the existence of biologically significant subclasses in the normal human colon.
Assuntos
Anticorpos Monoclonais , Colo/análise , Mucinas/imunologia , Adsorção , Especificidade de Anticorpos , Sequência de Carboidratos , Cromatografia DEAE-Celulose , Epitopos/análise , Humanos , Oligossacarídeos/análise , Peptídeo Hidrolases/metabolismo , Radioimunoensaio , Relação Estrutura-AtividadeRESUMO
We studied glycoprotein content of human colonic goblet cells, using a library of monoclonal antibodies (MAbs) directed against purified human colonic mucin (HCM). Using indirect immunofluorescence (IIF), we found that 17 of 23 anti-HCM MAbs stained some or all goblet cells of normal human colonic mucosa. We observed a variety of cellular staining patterns, including (a) diffuse (homogeneous) staining of intracellular mucin, (b) speckled (inhomogeneous) staining of mucin droplets, (c) peripheral staining of intracellular droplets, (d) cytoplasmic staining of goblet cells, and (e) apical (luminal) surface staining. Staining patterns were not associated with particular HCM species. In addition to variable patterns of IIF within individual cells, anti-HCM MAbs varied in the proportion of goblet cells stained. Some MAbs stained all goblet cells, while others stained a limited number of goblet cells. Although each goblet cell contained more than one type mucin, HCM species III, and IV and V appeared to exist in mutually exclusive goblet cell populations and it was possible to define at least seven subpopulations of goblet cells in colonic mucosa by their content of various combinations of HCM species. Anti-HCM MAbs stained goblet cells from other sites within the gastrointestinal tract to a varying extent. Anti-HCM MAbs also showed extensive cross-reactivity with rodent, rabbit, and monkey colonic mucosa. However, several anti-HCM MAbs stained only human colonic mucosa. These data show that human colonic mucosa contains discrete subpopulations of goblet cells that produce distinctive combinations of specific mucin glycoprotein species.