RESUMEN
OBJECTIVES: Many African countries have reported fewer COVID-19 cases than countries elsewhere. By the end of 2020, Guinea-Bissau, West Africa, had <2500 PCR-confirmed cases corresponding to 0.1% of the â¼1.8 million national population. We assessed the prevalence of SARS-CoV-2 antibodies in urban Guinea-Bissau to help guide the pandemic response in Guinea-Bissau. STUDY DESIGN: Cross-sectional assessment of SARS-CoV-2 antibody in a cohort of staff at the Bandim Health Project. METHODS: We measured IgG antibodies using point-of-care rapid tests among 140 staff and associates at a biometric research field station in Bissau, the capital of Guinea-Bissau, during November 2020. RESULTS: Of 140 participants, 25 (18%) were IgG-positive. Among IgG-positives, 12 (48%) reported an episode of illness since the onset of the pandemic. Twenty-five (18%) participants had been PCR-tested between May and September; 7 (28%) had been PCR-positive. Four of these seven tested IgG-negative in the present study. Five participants reported that somebody had died in their house, corresponding crudely to an annual death rate of 4.5/1000 people; no death was attributed to COVID-19. Outdoor workers had a lower prevalence of IgG-positivity. CONCLUSIONS: In spite of the low official number of COVID-19 cases, our serosurvey found a high prevalence of IgG-positivity. Most IgG-positives had not been ill. The official number of PCR-confirmed COVID-19 cases has thus grossly underestimated the prevalence of COVID-19 during the pandemic. The observed overall mortality rate in households of Bandim Health Project employees was not higher than the official Guinean mortality rate of 9.6/1000 people.
Asunto(s)
COVID-19 , SARS-CoV-2 , Adulto , Estudios Transversales , Atención a la Salud , Guinea Bissau/epidemiología , HumanosRESUMEN
The incidence of reported infections of non-typhoid Salmonella is affected by biases inherent to passive laboratory surveillance, whereas analysis of blood sera may provide a less biased alternative to estimate the force of Salmonella transmission in humans. We developed a mathematical model that enabled a back-calculation of the annual seroincidence of Salmonella based on measurements of specific antibodies. The aim of the present study was to determine the seroincidence in two convenience samples from 2012 (Danish blood donors, n = 500, and pregnant women, n = 637) and a community-based sample of healthy individuals from 2006 to 2007 (n = 1780). The lowest antibody levels were measured in the samples from the community cohort and the highest in pregnant women. The annual Salmonella seroincidences were 319 infections/1000 pregnant women [90% credibility interval (CrI) 210-441], 182/1000 in blood donors (90% CrI 85-298) and 77/1000 in the community cohort (90% CrI 45-114). Although the differences between study populations decreased when accounting for different age distributions the estimates depend on the study population. It is important to be aware of this issue and define a certain population under surveillance in order to obtain consistent results in an application of serological measures for public health purposes.
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Infecciones por Salmonella/epidemiología , Salmonella/aislamiento & purificación , Pruebas Serológicas/métodos , Adolescente , Adulto , Anciano , Estudios de Cohortes , Dinamarca/epidemiología , Métodos Epidemiológicos , Femenino , Humanos , Incidencia , Masculino , Persona de Mediana Edad , Infecciones por Salmonella/microbiología , Estudios Seroepidemiológicos , Adulto JovenRESUMEN
An increase in pertussis has been observed in several countries over the last decades, especially in adult populations. The seroprevalence of pertussis was determined in a cross-sectional study of the adult population in the Copenhagen area, Denmark, conducted between 2006 and 2008. Specific IgG antibodies against pertussis toxin (PT) were measured in 3440 persons resulting in an age-standardized seroprevalence of 3.0% (95% confidence interval 1.9-4.7) using an IgG anti-PT cut-off of 75 IU/ml. By using antibody decay profiles from longitudinal data the estimated seroincidence was 143/1000 person-years. In contrast, an incidence of 0.03/1000 person-years was estimated from the official data of notified cases during the same period. Of the investigated risk factors, only age and education were significantly associated with pertussis infection. This study indicates that pertussis is highly underestimated in the adult population in Denmark, which has implications for future prevention strategies, including raising the awareness of pertussis.
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Anticuerpos Antibacterianos/sangre , Bordetella pertussis/inmunología , Tos Ferina/epidemiología , Adolescente , Adulto , Anciano , Estudios Transversales , Dinamarca/epidemiología , Femenino , Humanos , Inmunoglobulina G/sangre , Incidencia , Masculino , Persona de Mediana Edad , Factores de Riesgo , Estudios Seroepidemiológicos , Tos Ferina/inmunología , Adulto JovenRESUMEN
The detection of urinary antigen is the most widely used method to diagnose Legionnaires' disease (LD), so it is important that these assays have a high sensitivity for the disease. In this study, we compare two kits for their ability to detect urinary antigen in urine samples from patients infected with Legionella species and L. pneumophila sero- and subgroups not considered as the most common causes of LD. Urine samples (n = 33) from 30 culture-proven cases of L. pneumophila serogroup (sg) 1, subgroup non-Pontiac infection, and urine samples (n = 35) from 32 cases of non-L. pneumophila species or non-sg 1 infection were examined using the Binax EIA and Biotest EIA kits. For both groups, the overall diagnostic sensitivity of the Binax kit was significantly better than the sensitivity of the Biotest kits (P < 0.0001). For the non-Pontiac group, the sensitivity was 81.8 and 42.4%, respectively, and for the non-sg1 group, it was 51.4 and 28.6%, respectively. It was concluded that the Binax kit was more suitable for the general diagnosis of LD than the Biotest kit, but we still need urinary antigen detection methods with higher sensitivity for non-sg1 LD.
Asunto(s)
Antígenos Bacterianos/análisis , Legionella/aislamiento & purificación , Legionelosis/diagnóstico , Juego de Reactivos para Diagnóstico , Orina/química , Orina/microbiología , Humanos , Técnicas para Inmunoenzimas/métodos , Sensibilidad y EspecificidadRESUMEN
Some anti-diabetic therapies can have adverse effects on bone health and increase fracture risk. In this study, we tested the skeletal effects of chronic administration of two Glucagon-like peptide-1 receptor agonists (GLP-1RA), increasingly used for type 2 diabetes treatment, in a model of osteoporosis associated bone loss and examined the expression and activation of GLP-1R in bone cells. Mice were ovariectomised (OVX) to induce bone loss and four weeks later they were treated with Liraglutide (LIR) 0.3mg/kg/day, Exenatide (Ex-4) 10 µg/kg/day or saline for four weeks. Mice were injected with calcein and alizarin red prior to euthanasia, to label bone-mineralising surfaces. Tibial micro-architecture was determined by micro-CT and bone formation and resorption parameters measured by histomorphometric analysis. Serum was collected to measure calcitonin and sclerostin levels, inhibitors of bone resorption and formation, respectively. GLP-1R mRNA and protein expression were evaluated in the bone, bone marrow and bone cells using RT-PCR and immunohistochemistry. Primary osteoclasts and osteoblasts were cultured to evaluate the effect of GLP-1RA on bone resorption and formation in vitro. GLP-1RA significantly increased trabecular bone mass, connectivity and structure parameters but had no effect on cortical bone. There was no effect of GLP-1RA on bone formation in vivo but an increase in osteoclast number and osteoclast surfaces was observed with Ex-4. GLP-1R was expressed in bone marrow cells, primary osteoclasts and osteoblasts and in late osteocytic cell line. Both Ex-4 and LIR stimulated osteoclastic differentiation in vitro but slightly reduced the area resorbed per osteoclast. They had no effect on bone nodule formation in vitro. Serum calcitonin levels were increased and sclerostin levels decreased by Ex-4 but not by LIR. Thus, GLP-1RA can have beneficial effects on bone and the expression of GLP-1R in bone cells may imply that these effects are exerted directly on the tissue.
Asunto(s)
Huesos/efectos de los fármacos , Receptor del Péptido 1 Similar al Glucagón/agonistas , Liraglutida/administración & dosificación , Osteogénesis/efectos de los fármacos , Osteoporosis/tratamiento farmacológico , Péptidos/administración & dosificación , Ponzoñas/administración & dosificación , Proteínas Adaptadoras Transductoras de Señales , Animales , Resorción Ósea , Huesos/diagnóstico por imagen , Huesos/metabolismo , Calcitonina/sangre , Diabetes Mellitus Tipo 2/tratamiento farmacológico , Modelos Animales de Enfermedad , Exenatida , Femenino , Glicoproteínas/sangre , Glicoproteínas/metabolismo , Inmunohistoquímica , Péptidos y Proteínas de Señalización Intercelular , Ratones , Ratones Endogámicos C57BL , Osteoblastos/citología , Osteoblastos/metabolismo , Osteoclastos/metabolismo , Osteocitos/citología , Ovariectomía , ARN Mensajero/metabolismo , Tibia/diagnóstico por imagen , Microtomografía por Rayos XRESUMEN
Mechanical failure of the fibrous cap of a vulnerable atherosclerotic plaque may lead to sudden plaque rupture and thus precipitate arterial thrombosis. Because ultrasound correlates strongly with mechanical features of tissues it might provide information on the stability of fibrous caps. The acoustic properties of the normal vessel wall and plaques, particularly fibrous caps of lipid-rich plaques, were evaluated in the aortic roots of six normal C57BL mice and 12 atherosclerosis-prone apoE-deficient (apoE(-/-)) mice by scanning acoustic microscopy (SAM). After processing, the attenuation of high-frequency (1.1 GHz) focused ultrasound was measured in unstained tissue sections by SAM followed by quantification of the amount and type of collagen in picrosirius red stained sections by means of polarized light microscopy (PLM). The acoustic and optical images were superimposed and ultrasonic attenuation was measured in different tissue components. Pertinent plaque features, particularly fibrous caps covering lipid-rich pools, were clearly visualized by both SAM and PLM. Collagen appeared green in thin fibrous caps and bright orange in thick caps by PLM. The attenuation of ultrasound was significantly higher in the collagen fibers with orange color compared to those with green color (17.2 versus 6.6 x 10(3) dB/mm). SAM has shown the possibility to characterize the types of collagen by high frequency intravascular ultrasound in vivo and it might improve our understanding of the vulnerable plaque and its sudden rupture from micro-mechanical point of view.
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Aorta/ultraestructura , Apolipoproteínas E/deficiencia , Arteriosclerosis/patología , Colágeno/ultraestructura , Metabolismo de los Lípidos , Animales , Aorta/metabolismo , Aorta/patología , Aorta/fisiopatología , Apolipoproteínas E/genética , Arteriosclerosis/metabolismo , Arteriosclerosis/fisiopatología , Fenómenos Biomecánicos , Ratones , Ratones Endogámicos C57BL , Microscopía/métodos , Microscopía de Polarización , UltrasonidoRESUMEN
The aims of the study were to evaluate characteristics of spontaneous motility and of the ascending excitatory peristaltic reflex (AEPR) and intraluminal cross-sectional area in the isolated perfused porcine duodenum. The parameters were measured by an intraluminal catheter by use of the perfused side-hole technique and impedance planimetry. Respiratory parameters such as pH and oxygen consumption and the arterial perfusion pressure were monitored and did not vary significantly throughout the study time. Spontaneous motility was intense at the beginning but declined and disappeared within 45-90 min. It was abolished by atropine, epinephrine, and UK-14,304 (an alpha 2-adrenoceptor agonist). Secondary motility was evoked by intraluminal balloon distensions by raising the balloon pressure to 1.5 kPa for 1-min periods. Reproducible results regarding the AEPR, external balloon diameters to elicit the AEPR, and intraluminal cross-sectional area were obtained. The order of potency (pD2 values) for inhibition of the AEPR was the selective M3-receptor antagonist 4-DAMP greater than atropine greater than the selective M2-receptor antagonist AFDX-116 greater than the selective M1-receptor antagonist pirenzepine greater than hexamethonium. 4-DAMP was 16 and 29 times more potent than AFDX-116 (P less than 0.02) and pirenzepine (P less than 0.02). None of the drugs altered the intraluminal cross-sectional area during the balloon distensions. The model provides the opportunity for physiological and pharmacological studies of duodenal motility and duodenal cross-sectional area devoid of extrinsic neural and endocrine effects. The abolishment of the AEPR by atropine is caused by blockade of the M3-receptor in the porcine duodenum.
Asunto(s)
Duodeno/fisiología , Motilidad Gastrointestinal/fisiología , Músculo Liso/fisiología , Animales , Análisis de los Gases de la Sangre , Duodeno/anatomía & histología , Duodeno/efectos de los fármacos , Estimulación Eléctrica , Electrofisiología , Motilidad Gastrointestinal/efectos de los fármacos , Técnicas In Vitro , Músculo Liso/efectos de los fármacos , Consumo de Oxígeno/efectos de los fármacos , Consumo de Oxígeno/fisiología , Parasimpatolíticos/farmacología , Perfusión , Peristaltismo/efectos de los fármacos , PorcinosRESUMEN
The influence of neurotransmitters on gastrointestinal motility is different in different segments of the gastrointestinal tract. To clarify the regulation of duodenal motility, the aim of the present study was to investigate the effects of alpha-adrenoceptor agonism and blockade and of galanin on duodenal motility. The study was undertaken in the isolated perfused porcine pancreatico-duodenal block. The agents under investigation were administered arterially. Duodenal motility was measured by means of a low-compliance perfusion system using an intraluminal catheter. In addition the concentration of galanin was measured in the portal effluent. We found that spontaneous motility was abolished by noradrenaline by an effect that was counteracted by the alpha 2-adrenoceptor antagonist idazoxan. In contrast, the selective alpha 1-adrenoceptor antagonist prazosin did not influence the effect of noradrenaline. Galanin, like noradrenaline, abolished duodenal motility. Furthermore, the concentration of galanin in the portal effluent was decreased by noradrenaline by an alpha 2-adrenoceptor mediated mechanism. We conclude that alpha 2-adrenoceptor activation and galanin inhibit duodenal motility and that the release of galanin from the pancreatico-duodenal preparation is reduced by alpha 2-adrenoceptors.
Asunto(s)
Duodeno/efectos de los fármacos , Motilidad Gastrointestinal/efectos de los fármacos , Norepinefrina/farmacología , Péptidos/farmacología , Antagonistas Adrenérgicos alfa/farmacología , Animales , Dioxanos/farmacología , Duodeno/fisiología , Galanina , Motilidad Gastrointestinal/fisiología , Idazoxan , Técnicas In Vitro , Neuropéptidos/farmacología , Neuropéptidos/fisiología , Norepinefrina/fisiología , Páncreas/efectos de los fármacos , Páncreas/fisiología , Péptidos/fisiología , Perfusión , Prazosina/farmacología , Receptores Adrenérgicos alfa/efectos de los fármacos , Receptores Adrenérgicos alfa/fisiología , PorcinosRESUMEN
The propagation speed (C) of surface acoustic waves (SAW), e.g. Rayleigh (R-waves) and longitudinal lateral waves (L-waves), the latter being the surface manifestation of the longitudinal waves, strongly reflect mechanical properties of materials. In view of an increasing interest in ultrasonic methodology in the field of bone biomechanics, we tested the hypothesis that both R- and L-waves can be excited in trabecular bone using an acoustic microscope at 1 GHz and that their speeds (C(R) and C(L)) can be extracted from V(z)-curves, i.e. plots of lens output voltage as a function of the lens focal point position with respect to the specimen surface. In accordance with V(z)-curves theoretically synthesized on the basis of incident field theory, experimental curves for canine femoral trabecular bone showed evidence of both R- and L-waves in almost all regions of recording. The measured CR ranged between 1.93 and 2.07 km/s (mean +/- SD.: 2.00=0.06 km/s) and the C(L) ranged between 2.33 and 4.33 km/s (3.37+/-0.61 km/s). Knowledge of both speeds allowed computation of a number of material constants by means of simple theory of elasticity and assumptions of the material density. We found values of Poisson ratio (v) ranging from 0.14 to 0.32 (0.23+/-0.07). Young's modulus (E) from 15 to 22.8 GPa (19.9+/-2.5 GPa) and the shear modulus (G) from 7.6 to 8.9 GPa (8.4+/-0.5 GPa). Anisotropy in the trabecular bone material was clearly detected at the micrometer level. In conclusion, the V(z)-curve method was successfully used to determine the distribution of material elastic constants of trabecular bone with micrometer resolution.
Asunto(s)
Huesos/fisiología , Microscopía/métodos , Acústica , Animales , Anisotropía , Fenómenos Biomecánicos , Perros , Elasticidad , Microscopía/instrumentación , Modelos BiológicosRESUMEN
The aim was to develop a method for the quantification of the stress-strain distribution of tubular organs in vivo using the porcine duodenum as an experimental model. We placed four electrodes for impedance planimetry and a 20 MHz ultrasound transducer inside an inflatable balloon mounted on a 6 mm-diameter probe for intraluminal use. By means of the ultrasound transducer and the impedance planimetric system, we measured the wall thickness and luminal cross-sectional area of the duodenum. We calculated the luminal radius on the basis of the latter. We validated ultrasonic measurements of wall thicknesses by comparing the former to microscopic measurements of the wall thickness of unstrained duodenal specimens in vitro. Also, we tested whether the magnitude of the applied balloon pressure affected ultrasonic measurements in vitro. The luminal and the outer radius increased non-linearly by a factor of three, rising steeply at low balloon pressures, moderately at higher pressures. The wall thickness decreased as an almost linear function of the applied balloon pressure, resulting in a 50% decrease. The stress-strain distribution calculated on the basis of the balloon pressures, the wall thicknesses and the luminal radii were non-linear. The wall reached a maximum circumferential strain of 1.71 at 6 kPa, corresponding to a stress of 64.8 kPa. Our system enabled us to quantify the stress-strain distribution of the porcine duodenum in vivo, and it may become a valuable tool for future biomechanical investigations of tubular organs in health and disease.
Asunto(s)
Duodeno/fisiología , Impedancia Eléctrica , Ultrasonografía Intervencional , Animales , Fenómenos Biomecánicos , Cateterismo/instrumentación , Modelos Animales de Enfermedad , Duodeno/anatomía & histología , Duodeno/diagnóstico por imagen , Elasticidad , Diseño de Equipo , Procesamiento de Imagen Asistido por Computador/instrumentación , Presión , Porcinos , Ultrasonografía Intervencional/instrumentaciónRESUMEN
The aim of the study is to devise a method for direct measurements of pressure, cross-sectional area (CSA) and wall thickness of a vessel when pressurised. Segments of porcine descending thoracic aorta from 22 and 35 kg pigs (in groups 1 and 2, respectively) and of abdominal aorta from 35 kg pigs are studied in vitro. Impedance planimetry provides measurements of the luminal pressure and CSA of the aorta. The wall thickness is measured simultaneously by means of 20 MHz A-mode ultrasound. The pressure, CSA and wall thickness are registered at different pressure at steady state. At maximum pressure, the CSAs are 107 +/- 10, 215 +/- 19 and 257 +/- 17 mm2 in the abdominal, group 1 and 2 segments, respectively. There is a difference between the abdominal group and groups 1 and 2 (p < 0.05), but not between group 1 and 2 segments (p > 0.2). At maximum pressure, the wall thickness is 0.52 +/- 0.04, 0.79 +/- 0.07 and 1.07 +/- 0.02 mm, in the abdominal group 1 and 2 segments, respectively (p < 0.05). The circumferential stress-strain relationship accorded well with stress = a x exp(b x strain). The stress-strain curve of the abdominal aortas is shifted to the left, indicating an increase (p < 0.05).
Asunto(s)
Aorta/fisiología , Animales , Aorta/anatomía & histología , Aorta/diagnóstico por imagen , Fenómenos Biomecánicos , Técnicas de Cultivo , Estrés Mecánico , Porcinos , UltrasonografíaRESUMEN
Acoustic microscopes can be used to measure Rayleigh and longitudinal or P-wave speeds in a specimen at microscopic resolution. The wave speeds are obtained from the interference pattern as a function of the defocus distance or V(z) curve. The received signal voltage amplitude Vis generated by two beams--the normally reflected central beam and a non-specularly reflected beam that strikes the fluid-solid interface at critical angle. It is shown in this paper that instead of analyzing the interference pattern between these two beams if we consider two other beams that follow the same path but travel through the coupling fluid multiple times before interfering then the V(z) curve generated by this higher order interference gives more accurate values for the material properties. The spacing distance between two successive dips of the V(z) curve is smaller for the higher order interference. The higher order interference, although weaker, gives more accurate results. Justification for the greater accuracy of the higher order interference is given in the paper. Material properties of silicon and bone are obtained by the new technique. Bones are microscopically heterogeneous and anisotropic. Anisotropic properties of homogeneous specimens can be obtained by the line focus acoustic microscope; however, it does not work when the specimen is microscopically heterogeneous. An attempt has been made here to obtain anisotropic properties of bones using point focus lens.
Asunto(s)
Huesos/fisiología , Silicio , Ultrasonido , Anisotropía , Elasticidad , Modelos TeóricosRESUMEN
The aim of this study was to evaluate the performance of an in-house ELISA for the diagnosis of Legionnaires' disease (LD) by detection of IgM and IgG antibodies against Legionella (L.) pneumophila serogroups (sg) 1, 3 and 6. The evaluation was done throughout a two-year period in a diagnostic routine laboratory. Furthermore, the sensitivity of four different methods, the in-house L. pneumophila antibody test (ELISA), the urinary antigen test (Binax® EIA), an in-house PCR and culture, both alone and in combination was evaluated. From 2008 to 2010, 12,158 serum samples from 10,503 patients were analysed. During the same period, 361 cases of laboratory-confirmed LD cases were recorded in Denmark, but of these only 113 had a serum sample examined. The positive predictive value of the in-house ELISA was calculated to be 12.8 and the negative predictive value was 99.6, using only the confirmed LD cases as true positives. The sensitivity of the in-house ELISA for the detection of IgM and IgG antibodies in the confirmed LD cases was 61% and 36%, respectively. By combining the two ELISA assays the sensitivity increased to 66%. The sensitivity of the Legionella urinary antigen test (Binax® EIA) was 63%, of the in-house PCR 87% and of culture 69%. When all the different methods were combined, a higher sensitivity was calculated--for in-house ELISA (IgM+IgG) and Binax® EIA 91%, in-house ELISA (IgM+IgG) and in-house PCR 93%, in-house ELISA (IgM+IgG) and culture 93%, Binax® EIA and in-house PCR 79%, Binax® EIA and culture 68% and in-house PCR and culture 94%. This study confirms that the detection of IgG and IgM antibodies by ELISA is an important diagnostic tool, also during the initial phase of the disease. Furthermore, we showed that LD in Denmark with or without serum samples collected exhibits the same age and sex distribution and epidemiology, as in the rest of Europe, i.e., mostly men are infected, infections are mostly community acquired, followed by infection from travelling abroad. Apart from patients with notified LD, the patients investigated by serology were evenly distributed in all age groups; there was only a slightly higher ratio of men tested for "atypical pneumonia" in the serology laboratory.
Asunto(s)
Anticuerpos Antibacterianos/sangre , Pruebas Diagnósticas de Rutina/métodos , Ensayo de Inmunoadsorción Enzimática/métodos , Inmunoglobulina G/sangre , Inmunoglobulina M/sangre , Legionella pneumophila/inmunología , Enfermedad de los Legionarios/diagnóstico , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Anticuerpos Antibacterianos/inmunología , Niño , Femenino , Humanos , Inmunoglobulina G/inmunología , Inmunoglobulina M/inmunología , Legionella pneumophila/clasificación , Legionella pneumophila/genética , Legionella pneumophila/aislamiento & purificación , Enfermedad de los Legionarios/inmunología , Enfermedad de los Legionarios/microbiología , Masculino , Persona de Mediana Edad , Reacción en Cadena de la Polimerasa , Adulto JovenAsunto(s)
Momias/patología , Entierro/historia , Dinamarca , Medicina Legal , Historia del Siglo XX , Historia Medieval , Humanos , Intestinos/patología , Imagen por Resonancia Magnética , Masculino , Momias/diagnóstico por imagen , Momias/historia , Paleopatología , Cráneo/diagnóstico por imagen , Tomografía Computarizada por Rayos XRESUMEN
The interaction of calreticulin with native and denatured forms and polypeptides in proteolytic digests of proteins representing structural classes of all-alpha-helix (hemoglobin, serum albumin), all-beta-sheet (IgG) and alpha-helix + beta-sheets (lysozyme, ovalbumin) was investigated. The binding of calreticulin to denatured proteins was found to depend on conformation and structural class of the protein. No interaction was observed with the native proteins, whereas binding was seen for the denatured proteins, the order of interaction being lysozyme = IgG > ovalbumin >> hemoglobin = serum albumin. Moreover, the interaction between calreticulin and the heat-denatured proteins depended on the temperature and time used for denaturation and the degree of proteolytic fragmentation. Calreticulin bound well to peptides in proteolytic digests from protease K or chymotrypsin treatment of lysozyme, IgG and ovalbumin but weakly or not at all to peptides in proteolytic digests of hemoglobin and serum albumin. Synthetic peptides from lysozyme and ovalbumin confirmed binding to hydrophobic peptides from these proteins. These results show that calreticulin has the ability to interact with denatured and fragmented forms of proteins with a preference for beta-strand structure and hydrophobicity.
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Calreticulina/metabolismo , Péptidos/metabolismo , Proteínas/metabolismo , Calreticulina/química , Hemoglobinas/química , Hemoglobinas/metabolismo , Humanos , Inmunoglobulina G/química , Inmunoglobulina G/metabolismo , Muramidasa/química , Muramidasa/metabolismo , Ovalbúmina/química , Ovalbúmina/metabolismo , Péptidos/química , Proteínas Gestacionales/química , Proteínas Gestacionales/metabolismo , Unión Proteica , Desnaturalización Proteica , Pliegue de Proteína , Dominios y Motivos de Interacción de Proteínas , Estructura Secundaria de Proteína , Proteínas/química , Albúmina Sérica/química , Albúmina Sérica/metabolismo , TemperaturaRESUMEN
Four commercially available kits from (1) Focus Diagnostics, (2) SERION, (3) Zeus and (4) Vircell for detection of antibodies to Legionella pneumophila were evaluated with panels of sera from patients with proven Legionella infection (n = 81) and/or other bacterial infections (n = 75). An in-house indirect Legionella immunofluorescence antibody test (IF test) was used as reference. All sera from the laboratory-proven Legionella pneumophila cases [culture, urinary antigen test and/or polymerase chain reaction] of Legionella infection were found to be positive by the in-house IF test. The relative sensitivity for Focus Diagnostics, SERION, Zeus and Vircell kits was 81.5, 76.5, 68.8 and 62.5%, respectively, and the false-positive rate was 16.0, 5.6, 29.0 and 2.7%, respectively. The in-house IF test had a false-positive rate of 4.0%. It was found that none of the four commercial kits were as sensitive and specific as the in-house IF test.
Asunto(s)
Anticuerpos Antibacterianos/sangre , Técnica del Anticuerpo Fluorescente Indirecta/métodos , Legionella pneumophila/inmunología , Enfermedad de los Legionarios/diagnóstico , Juego de Reactivos para Diagnóstico , Reacciones Falso Positivas , Humanos , Sensibilidad y EspecificidadRESUMEN
OBJECTIVE: Gc globulin (vitamin D-binding protein) is a component of the extracellular actin scavenger system. The level of Gc globulin is reduced in patients with fulminant hepatic failure, septic shock and trauma. Furthermore, low levels of Gc globulin in patients with fulminant hepatic failure and multiple trauma have been found to correlate with the morbidity and mortality of patients. Owing to a large increase in the turnover of Gc globulin upon complex formation with actin, it may be important to determine both the total Gc globulin concentration and the degree of complexing with actin for estimating the clinical prognosis of a patient. For this reason, we have compared a crossed immuno-electrophoresis method (CIE), suitable for visualizing the degree of complexing with actin, with a rocket immuno-electrophoresis method (RIE), previously used for determination of the complex degree. MATERIAL AND METHODS: Sera from healthy donors and from patients with acetaminophen-induced liver disease or trauma were investigated using CIE, RIE and enzyme-linked immunosorbent assay (ELISA). RESULTS: Using the CIE, no Gc globulin-actin complexes were detected among healthy donors. Complexes were present in 21 of 39 patients with liver disease and 3 of 37 trauma patients. High complex ratios (> 20 %) were found in 6 of 7 patients with hepatic encephalopathy. Using the RIE, complexes were detected in most samples. CONCLUSION: The results show that the CIE method may be used for determining the degree of actin complexing in conjunction with ELISA or RIE in determining the levels of total Gc globulin.
Asunto(s)
Actinas/sangre , Inmunoelectroforesis Bidimensional/métodos , Inmunoelectroforesis/métodos , Proteína de Unión a Vitamina D/sangre , Acetaminofén/efectos adversos , Actinas/metabolismo , Calibración , Enfermedad Hepática Inducida por Sustancias y Drogas , Ensayo de Inmunoadsorción Enzimática/métodos , Gelsolina/química , Encefalopatía Hepática/sangre , Encefalopatía Hepática/inducido químicamente , Humanos , Hepatopatías/sangre , Unión Proteica , Reproducibilidad de los Resultados , Temperatura , Proteína de Unión a Vitamina D/metabolismo , Heridas y Lesiones/sangreRESUMEN
Calreticulin has been reported to be an autoantigen in various autoimmune connective tissue diseases and in coeliac disease. Previous studies have used incubation buffers with low salt and low detergent concentrations (low stringency conditions) with serum albumin or other proteins as a blocking agent. Using these conditions we found a relatively high level of non-specific binding in many sera. Antibodies to proteins that are used as blocking reagents in ELISA (bovine serum albumin (BSA), ovalbumin, skimmed milk powder) are frequently present in sera, and these may cause false-positive results. Moreover, the low isoelectric point of calreticulin and its chaperone properties may give rise to false-positive results under low stringency conditions. We report that the use of a simple buffer without protein (50 mM Tris, pH 7.5, 1% Tween 20, 0.3 M NaCl) removes most of the problems with unwanted binding (high stringency conditions). Using the high stringency conditions, we screened sera from 107 patients with systemic lupus erythematosus, sera from patients with other systemic autoimmune diseases and from children with coeliac disease for the presence of high-affinity calreticulin autoantibodies by immunoblotting and ELISA. None of the sera contained high-affinity calreticulin antibodies. It is concluded that calreticulin is not a common autoantigen in patients with autoimmune connective tissue diseases or coeliac disease.
Asunto(s)
Autoanticuerpos/sangre , Calreticulina/inmunología , Enfermedad Celíaca/inmunología , Enfermedades Reumáticas/inmunología , Ensayo de Inmunoadsorción Enzimática , Humanos , ImmunoblottingRESUMEN
OBJECTIVES: Determination of physical and mechanical characteristics and properties of thin-walled (0.39 mm) expanded polytetrafluoroethylene stretch vascular prostheses by time-resolved scanning acoustic microscopy (TR-SAM). MATERIALS AND METHODS: Samples of Stretch Gore-Tex were mounted on polymethylmethaacrylate substrate, and ethyl alcohol was used as acoustic couplant. A 10 MHz ultrasound transducer mounted on a SAM50 scanning acoustic microscope generated short, pulsed sound waves. Reflected signals were used for imaging and for computer analysis of time resolved wave forms acquired by a digital sampling oscilloscope to calculate physical and mechanical characteristics of the material. RESULTS: Graft wall thickness: 0.395 (0.352/0.401) mm; graft wall sound wave penetration velocity: 1111 (1083/1129) ms-1; acoustic impedance: 3.685 (2.975/4.370) 10(6) kg m-3s-1; attenuation coefficient: 0.144 (0.096/0.229) dB mm-1 MHz-1; material density: 3360 (2712/3982) kg m-3; elastic stiffness: 4.06 (3.35/4.83) GPa (median and (25/75) percentiles; n = 8 samples, 10 measurements spaced 100 microns apart in each sample). CONCLUSIONS: TR-SAM can effectively quantify ePTFE graft wall properties, and new data on graft properties have been provided. The acoustic impedance and elastic stiffness of the ePTFE graft wall are 2.2 and 1.6 times higher, respectively, than in human coronary arteries, and it is highly reflective of ultrasound.