Performance evaluation of density measurements of axial and peripheral bone with x-ray and gamma-ray computed tomography

We examined sources of error in bone measurements made with computed tomography (CT) using a whole-body scanner (GE 8800) and a peripheral-bone CT scanner (developed at the University of Alberta). We investigated the influence of various factors on trabecular bone density: homogeneity and noise in the image plane, linearity of calibration, body size, effects of cortical bone, and the image analysis procedure. With the GE 8800 scanner, the precision (SD) of measurements of a single vertebra is expected to be +/- 1.65% (noise: +/- 0.22%, calibration: +/- 1.3%, analysis: +/- 1%); the accuracy, excluding consideration of marrow fat, varied between -2.7 and +7.3% (compact-bone thickness: 2-5%, body size: -2.5 - +1.5%, calibration: -0.47 - +0.77%). With the peripheral-bone CT scanner, the total precision error (+/- 0.53%) was dominated by noise, with only a minor contribution from the analysis procedure (+/- 0.04%); accuracy varied between -0.6 and +3.4% (effect of cortical bone: up to 3.0%; changes in size of object: -0.59 - +0.4%). The magnitude of these errors was determined under 'ideal' conditions, mostly through phantom measurements; therefore, the errors represent optimistic lower limits in clinical application. Furthermore, measurements of density of cortical bone were not reliable for bone thicknesses of less than about 4 mm with the GE 8800 scanner and less than about 1.5 mm with the peripheral scanner.     

Morphological characteristics of clinically significant coronary artery stenosis in stable angina

All segments of clinically significant stenosis in the coronary arteries of 54 men with stable angina were categorised according to the position of the plaques (eccentric or concentric) and the presence or absence of a pool of extracellular lipid. In the group as a whole, stenosis of greater than 50% by diameter was caused by concentric fibrous plaques in 48% of lesions, by concentric lipid plaques in 28%, by eccentric fibrous plaques in 12%, and by eccentric lipid plaques in 12%. In addition, 43 of the 54 patients had one or more stenoses with multiple channels (recanalisation). Eccentric plaques with an arc of normal vessel wall occupying more than 16% of the circumference of the residual lumen were considered to have a vasospastic potential and made up 15% of all lesions with stenosis of greater than 50% by diameter. Forty four per cent of plaques causing stenosis between 30% and 50% by diameter were eccentric and retained a considerable arc of normal media. These lesions were often in series with segments of higher grade stenosis that did not have an arc of normal media. The overall frequency of plaque types gave no indication of the proportions of different plaque types within an individual. In 15% of patients all the plaques causing greater than 50% diameter stenosis were fibrous and in 13% of patients all the plaques were of the lipid type. Most patients had mixtures of all plaque types in varying proportions. Plaques with a large pool of lipid were not found in 33% of patients whereas they formed greater than 90% of the plaques in 9% of patients. No segments of stenosis > 50% by diameter with a vasospastic potential were found in 44% of the patients but one or more such plaques was present in the the remaining 56%. Three patients (6%) each had five separate segments of stenosis with a vasospastic potential. The results indicate that even in a population of men with stable angina in whom diabetes is excluded the distribution of types of atheromatous lesions is very heterogenous.     

LC-MS/MS测定人血浆中对乙酰氨基酚及其人体药动学和生物等效性研究

目的 建立一种快速、灵敏的高效液相色谱-串联质谱（HPLC-MS/MS）方法以测定人血浆中对乙酰氨基酚浓度,并应用于两种对乙酰氨基酚制剂的人体药代动力学和生物等效性研究。方法 以替硝唑为内标,200μL血浆样品经5倍于其体积的乙酸乙酯液液萃取,再经Waters XBridge? C18柱等度洗脱分离后导入串联质谱,以正离子多反应监测模式进行定量分析,对乙酰氨基酚和内标的选择性反应离子对分别是m/z 152→110和248→121。方法经验证后应用于19名健康受试者单剂量空腹口服两种对乙酰氨基酚制剂500mg后药代动力学和生物等效性的研究。结果 血浆中对乙酰氨基酚在0.1～8.0 μg·mL-1范围内线性良好（r2 > 0.99）,最低检测限为 0.1 μg·mL-1,提取回收率为91.0%～98.7%,日内和日间准确度分别为98.8%～111.3% （精密度：CV ? 9.03%）和94.9%～102.6% （精密度：CV ? 10.68%）。生物等效性试验中,受试制剂与参比制剂的主要药代动力学参数Cmax、AUC0-t和AUC0-∞ 几何均值比的90%置信区间分别为83.50%～105.79%,94.25%～101.54%和93.24%～101.02%,均落在生物等效可接受标准80.00%～125.00%范围内。结论 所建立测定人血浆中对乙酰氨基酚浓度的HPLC-MS/MS法具有快速灵敏、回收率高、选择性好的特点,适用于对乙酰氨基酚片人体药代动力学和生物等效性研究。受试制剂与参比制剂在人体内吸收速度和程度相似,两种制剂生物等效。     

Antiviral immune responses in gene-targeted mice expressing the immunoglobulin heavy chain of virus-neutralizing antibodies

Two gene-targeted immunoglobulin heavy chain transgenic mouse strains, TgH(KL25) and TgH(VI10), expressing neutralizing specificities for lymphocytic choriomeningitis virus and vesicular stomatitis virus, respectively, have been generated. Three days after lymphocytic choriomeningitis virus infection, TgH(KL25) mice showed a thymus-independent neutralizing IgM response followed by thymus-dependent (TD) IgG. In contrast, WT mice mounted only a TD IgG response around day 80. These observations indicated that not only structural properties of the virus but also immunological parameters such as the frequency of B cells were indicative for the induction of thymus-independent versus TD Ig responses. Na?ve vesicular stomatitis virusspecific Ig heavy chain transgenic mice displayed greatly elevated natural antibody titers. However, despite these high na?ve titers, de novo activation of na?ve CD4+ T and B cells was not blocked. Therefore, B cells giving rise to natural antibodies do not participate in virus-induced antibody responses.     

Adhesion of platelets to surface-bound fibrinogen under flow

After platelet activation, fibrinogen mediates platelet-platelet interactions leading to platelet aggregation. In addition, fibrinogen can also function as a cell adhesion molecule, providing a substratum for adhesion of platelets and endothelial cells. In this report, we studied the adhesion of platelets to surface-immobilized fibrinogen under flow in different shear rates. Heparinized whole blood containing mepacrine-labeled platelets was perfused for two minutes at various wall shear rates from 250 to 2,000 s-1 in a parallel plate flow chamber. The number of adherent fluorescent platelets was quantitated every 15 seconds with an epifluorescent videomicroscope and digital image processing system. When compared with platelet adhesion and aggregation seen on glass surfaces coated with type I bovine collagen, a significant increase in platelet adhesion was observed on immobilized fibrinogen up to wall shear rates of 800 s-1. The adherent platelets formed a single layer on fibrinogen-coated surfaces. Under identical conditions, no significant adhesion was observed on fibronectin- or vitronectin-coated surfaces. Although platelet adhesion to collagen was substantially inhibited by the platelet inhibitors prostaglandin E1 and theophylline, these inhibitors had no effect on platelet adhesion to fibrinogen. Platelets adhered to recombinant homodimeric wild-type (gamma 400-411) fibrinogen, but not to the recombinant homodimeric gamma' variant of fibrinogen. Platelet adhesion to recombinant fibrinogen with RGD to RGE mutations at positions alpha 95-97 and alpha 572-574 was similar to that with plasma-derived fibrinogen. These results show that platelets adhere to fibrinogen-coated surfaces under moderate wall shear rates, that the interaction is mediated by the fibrinogen 400-411 sequence at the carboxy-terminus of the gamma chain, and that the interaction is independent of platelet activation and the RGD sequences in the alpha chain.     

The development of cellular immunity to Epstein-Barr virus after allogeneic bone marrow transplantation

Epstein-Barr virus-induced lymphoproliferative disease (EBV-LPD) is a potentially lethal complication during the first 6 months after allogeneic bone marrow transplantation (BMT). To determine whether deficiencies of EBV-specific cellular immunity contribute to EBV-LPD susceptibility and distinguish patients at risk, we performed limiting dilution analysis to quantify anti-EBV cytotoxic T-lymphocyte precursor (CTLp) frequencies in 26 recipients of unmodified or T-cell-depleted (TCD) grafts from EBV-seropositive donors. At 3 months post-BMT (n = 26), only five patients had EBV CTLp frequencies in the range of seropositive normal controls, irrespective of the type of transplant administered. By 6 months post-BMT, 9 of 13 patients tested had EBV CTLp frequencies within the normal range. The time period in which these patients had deficient cellular immunity to EBV corresponds to the period in which we have observed EBV-LPD in most prior patients. One patient with a low EBV CTLp frequency at 4 months post-BMT developed an EBV-LPD. Within 2 weeks of receiving an infusion of donor peripheral blood mononuclear cells (PBMC) providing less than 1,200 EBV- specific cytotoxic T-cell precursors, populations of EBV-specific CTL in the circulation were restored to levels detected in normal seropositive adults. Concurrently, the patient achieved a regression of the EBV-LPD, which has been sustained without further therapy. These studies indicate that recipients of both unmodified and TCD marrow grafts have profound deficiencies of EBV-specific T cell-mediated immunity early posttransplant, and that the period of risk for EBV-LPD closely corresponds to this interval of severe deficiency. Treatment of one patient with EBV-LPD with marrow donor-derived PBMC induced a rapid expansion of EBV-specific cytotoxic T-cell populations that occurred contemporaneously with the clinical regression of disease.     

对产前母体苯巴比妥给药以预防针对新生儿和胎儿溶血病的婴儿换血

胎儿和新生儿溶血病(HDFN)可导致严重的围产期疾病,包括高胆红素血症及其引起的核黄疸。血红素分子解离为胆红素并产生免疫介导的溶血。它与白蛋白结合后转运到肝细胞,再由尿苷二磷酸葡萄糖醛酸转移酶(UGT)酶家族作用而葡萄糖醛酸化。对于人类,这种代谢途径中主要的酶是胆红素-