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91.
Migration of leukocytes across endothelium and beyond: molecules involved in the transmigration and fate of monocytes 总被引:13,自引:0,他引:13
Passage of leukocytes across the endothelial lining into sites of inflammation has been shown to be regulated largely by platelet/endothelial cell adhesion molecule-1 (PECAM/CD31). We summarize recent work from our laboratory documenting that PECAM is involved both in diapedesis and the subsequent step of migration across the basal lamina. The former process involves a homophilic interaction between the amino-terminal extracellular domains of PECAM on the leukocyte and on the endothelial cell. The latter process involves a heterophilic interaction between the membrane-proximal extracellular domain of PECAM and an unknown ligand(s) in the subendothelial basal lamina. These findings are demonstrated in both in vitro and in vivo models. For monocytes, however, transmigration is just the first step in the next phase of their lives. In addition to their specific recruitment during the inflammatory response, many monocytes constitutively leave the bloodstream to enter tissues. However, only a fraction of these become tissue macrophages. In an in vitro model of monocyte emigration, approximately half of the leukocytes that initially transmigrate an endothelial monolayer migrate back out in the basal-to-apical direction within the next 2 days. This reverse transmigration cannot be blocked by anti-PECAM reagents, but involves p-glycoprotein and tissue factor expressed on the leukocytes. The reverse transmigrating cells are phenotypically dendritic cells (DC). Their maturation to mature DC can be accelerated by inclusion of inflammatory stimuli in the co-culture. The cells that remain behind in the subendothelial collagen are phenotypically macrophages. We postulate that a major source of DC in lymph nodes is cells derived from monocytes that enter a tissue via the blood and leave several days later via afferent lymphatic channels. 相似文献
92.
Determination of the concentration of osteocalcin in rat serum is frequently performed using a commercially available radioimmunoassay
(RIA). However, this assay takes 3 days to complete, uses radioactive material, and has a narrow linear range. The limited
range of the RIA makes it necessary to test multiple dilutions of the sample which frequently results in values that differ,
depending on the dilution. In order to overcome these limitations, we have developed an ELISA that utilizes the same standards
and anti-rat osteocalcin antiserum, as is used in the RIA. The principle of the ELISA is that the osteocalcin in the sample
competes with osteocalcin previously immobilized on a microtiter plate to bind to the available anti-rat osteocalcin antibodies.
The amount of antibody bound to the immobilized osteocalcin is determined colorimetrically using a secondary antibody coupled
to alkaline phosphatase. This ELISA has a three-log linear response with a sensitivity of 0.1–0.15 ng/ml and intra- and interassay
coefficient of variance (CV) values of less than 10%. Most importantly, the assay is rapid and only requires a 2-hour incubation
of the sample with the antiserum. The incubation time is important since we and others have observed a significant decrease
in the osteocalcin level from serum samples incubated for long periods of time with the antiserum, presumably due to degradation
of the osteocalcin. In general, the commercially available RIA gives osteocalcin values that are one-half to one-fourth that
of the ELISA because the RIA requires a 48-hour incubation time.
Received: 14 November 1997 / Accepted: 9 July 1998 相似文献
93.
The sequence H236-256 of the measles virus (MV) hemagglutinin (H) contains the sequential epitope of the neutralizing and protective monoclonal antibody (mAb) BH129 with the minimal epitope E(245)L-QL(249). Using this mAb, we have recently developed 7mer mimotopes binding up to 135x better than the corresponding 7mer epitope H244-250. In this study, we combined T cell epitopes (TCE) with either highly crossreactive 7mer mimotopes, 13mer mimotopes or less crossreactive MV-derived B cell epitopes (BCE). Antigenicity of these TBB, TTB and TTBB peptides was determined with BH129 in a competition ELISA against MV. We found that chimeric peptides including mimotopes were up to 80x better binders to the mAb than peptides containing the original BCEs. All peptides irrespective of their antigenicity were used for immunization to compare their virus- crossreactive immunogenicity. Unexpectedly, none of the highly antigenic mimotope-based peptides induced MV-crossreactive antibodies. In contrast, a number of peptides with the viral BCE sequence that did not bind to the mAb, induced MV-crossreactive and even neutralizing antibodies.This report describes a striking example of disparity between antigenicity and crossreactive immunogenicity and casts considerable doubt on the predictive value of antigenicity in immunogenicity studies, considerably complicating the selection of potential vaccine candidates. 相似文献
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Objective Morbidity and mortality remain high amongst babies ventilated for a respiratory distress syndrome (RDS). Whether newly developed ventilators allowing high frequency ventilation such as high frequency flow interrupted ventilation (HFFIV) could decrease the morbidity and the mortality was investigated in a randomized study.Design Preterm babies weighing 1800g suffering from RDS and ventilated by conventional mechanical ventilation (CMV) were randomized to be further ventilated either by CMV (group CMV) or by HFFIV (group HFFIV) when peak inspiratory pressure (PIP) on CMV was 20cmH2O.Setting The study was undertaken in the neonatal intensive care unit of the Erasmus Hospital.Patients 24 patients entered into the investigation and were randomized but 2 patients were removed from the study because the switch over to HFFIV failed. Eight of the 12 CMV patients and 5 of the 10 HFFIV patients completed the study.Measurements and results Clinical variables, blood gas analysis and ventilatory variables were looked at. There were no differences in mortality, in incidence of air leaks and pulmonary complications or in blood gas analysis. Bronchopulmonary dysplasia was not decreased by the use of HFFIV.Conclusion It is concluded that HFFIV is safe although it offers no concrete advantages over CMV when applied as we did in a low pressure approach. 相似文献
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99.
Preliminary data from nonrandomized clinical trials suggest that, in selected subgroups of patients, the implantation of endovascular stents may improve the initial results of coronary dilatation, may successfully treat arterial dissection and abrupt closure, and may reduce the incidence of recurrent stenosis. The widespread use of stents remains limited, however, by the need for intensive anticoagulation to prevent arterial thrombosis. The development of sustained-release drug delivery systems and gene-transfer technology may enable local delivery of antithrombotic and antiproliferative therapies that would greatly increase the safety and applicability of these devices. 相似文献
100.
R M Quinlivan C R Muller M Davis N G Laing G A Evans J Dwyer J Dove A P Roberts C A Sewry 《Archives of disease in childhood》2003,88(12):1051-1055
Central core disease (CCD) is a dominantly inherited congenital myopathy allelic to malignant hyperthermia (MH) caused by mutations in the RYR1 gene on chromosome 19q13.1. Eleven individuals with RYR1 mutations are described. Four index cases showed features consistent with a congenital myopathy (hypotonia, delayed motor milestones, and skeletal abnormalities including congenital hip dislocation and scoliosis). All four cases and subsequently seven other family members were found to possess novel mutations in the RYR1 gene. The degree of disability varied from one clinically normal individual, to another who had never achieved independent ambulation (the only patient with a de novo mutation). Four cases showed a mild reduction in vital capacity, repeated nocturnal polysomnography showed hypoxaemia in one case. A variety of muscle biopsy features were found; central cores were absent in the youngest case, and the biopsy specimens from two others were more suggestive of mini-core myopathy. In all cases missense mutations in exons 101, 102, and 103 of the RYR1 gene on were found. Future laboratory diagnosis of suspected cases and family members will be less invasive and more accurate with DNA analysis. Clinicians, especially paediatricians and orthopaedic surgeons, should be aware of this disorder because of the potential risk of MH. 相似文献