CD24 expression on human keratinocytes

Abstract: CD24 or Nectadrin is a cell surface glycoprotein expressed in pre-B lymphocytes, T lymphocytes, neurons, muscle cells and carcinoma cells. Its function is not completely known, but it has been suggested that it is involved in cell adhesion and signalling. CD24 has recently been identified as the human molecule homologous to the murine heat-stable antigen (HSA). HSA is expressed by murine keratinocytes and delivers costimulatory signals in T-cell activation. Long-term cultures of normal human keratinocytes (HKC) were obtained from skin of human female breast sections and either left untreated or were treated with phorbol-12-myristate-13-acetate (PMA) at 10–100 ng/ml, calcium 0.5–2 mM or IFN-γ 100–1000 U/ml, for 24–48 h. Using RT-PCR and flow cytometry we showed that HKC express low levels of CD24 even under basal conditions, and the treatment with calcium, PMA or IFN-γ increased levels of CD24 mRNA and protein. To the best of our knowledge, this is the first report to measure CD24 expression and production by cultured HKC in basal conditions and after stimulation. Further studies are needed to determine biological and therapeutical relevance of these findings.     

Detection of HLA-A*24 null alleles by DNA typing methods

Abstract: A number of cases have been identified (seven unrelated individuals from the Northern Ireland bone marrow donor registry and two family groups) where an HLA-A*24 allele fails to express the normal HLA-A24 antigen. Family information has revealed common haplotypes with respect to each non-expressed allele indicating that the occurrence of these mutations has been a recent event. Two methods for the clinical typing of these alleles have been evaluated - PCR-SSOP and PCR-SSCP analysis.     

Ambulatory 24-hour esophageal pH monitoring

If 24-hour esophageal pH monitoring is to be a useful diagnostic tool, it must reliably discriminate gastroesophageal reflux patients despite daily variations in distal esophageal acid exposure. To address this issue, we studied 53 subjects (14 healthy normals, 14 esophagitis patients, and 25 patients with atypical symptoms) with two ambulatory pH tests performed within 10 days of each other. Intrasubject reproducibility of 12 pH parameters to discriminate the presence of abnormal acid reflux was determined. As a group, the parameters of percent time with pH<4 (total, upright, recumbent) were most reproducible (80%). Therefore, a subject was defined as having gastroesophageal reflux disease if at least one of these three values were abnormal. Intrasubject reproducibility for the diagnosis of reflux disease was 89% for the entire sample. Among subsets, the reproducibility was 93% for the normals and esophagitis patients and 84% for the atypical symptom patients. Total percent time with pH<4 was the single most discriminate pH parameter (85%) and nearly equaled that of the three combined parameters (89%). The intrasubject variability of this parameter was determined by the mean ±2sd of the relative differences between the two test results for all 53 subjects. Total percent time with pH<4 may vary between tests by a factor of 3.2-fold or less (218% higher to 69% lower). We conclude: (1) ambulatory 24-hr esophageal monitoring is a reproducible test for the diagnosis of gastroesophageal reflux disease; and (2) the large intrastudy variability in 24-hr total acid exposure may limit this test's usefulness as a measurement of therapeutic improvement.Supported, in part, by Public Health Services Grant AM 34200-01A1 from NIADDIK.     

基于非标准协议无线通信的个人健康监护系统设计

基于非标准协议的短距离无线通信，具有成本低、易开发、低功耗的优点，非常适用于小范围内的数据传输。提出一种基于非标准无线通信芯片nRF24L01的个人健康监护系统设计方案，并将其与现有的、基于标准协议无线通信的个人健康监护系统进行比较。结果表明：基于非标准协议无线通信的个人健康监护系统具有效率高、功耗低、通信性能好的特点。     

Impact of Ingested Liquids on 24-Hour Ambulatory pH Tests

A prospective investigation of the impact ofingested liquids on 24-hr pH test scores was conducted.Eighty-two patients contributed 142 samples. The liquidsused were coffee/tea (N = 35), water (N = 32), fruit juice (N = 29), cola (N = 34), and beer (N =12). The pH of cola, juice, and beer are approximately3.0. The parameters studied included: total test time,total drink time, total minutes of pH < 4.0 during drink, minutes of pH < 4.0 10 min beforedrink, and minutes of pH < 4.0 10 min followingdrink. Analysis was performed using one-way ANOVA andrepeated measures. Age of patients, total test time, and total time pH < 4.0 were notsignificantly different (P > 0.05). The total time toconsume the drink was significantly greater (P <0.05) for beer than all other liquids. The total time(7.7 ± 6.0 min) pH < 4.0 for cola wassignificantly different (P < 0.023) than beer (3.3± 3.7 min), tea/coffee (1.4 ± 6.5 min),and water (1.1 ± 2.5 min). The percentage oftotal time pH < 4.0 was not significantly different (P >0.05) among any of the liquids. The percentage of timepH < 4.0 during the drink was the highest for cola(63 ± 47%) and juice (51 ± 57%); water,coffee/tea, and beer were not significantly different (P> 0.05). Although the impact of cola and juice werethe greatest, none of these had an impact that exceeded0.5%. The lack of impact of beer appears to be due to the increased period of time it takes toconsume. We conclude that the impact of ingested fluidsis minimal and can probably be disregarded in mostpatient groups.     

Characteristics of β-endorphin-induced histamine release from rat serosal mast cells Comparison with neurotensin,dynorphin and compound 48/80

Summary Rat peritoneal mast cells were exposed to the neurohormone and basic opioid peptide 24/xxlarge946.gif" alt="beta" align="MIDDLE" BORDER="0">-endorphin. 24/xxlarge946.gif" alt="beta" align="MIDDLE" BORDER="0">-Endorphin induced a dose-dependent release of histamine from the mast cells. A significant histamine release was found at 5 24/xxlarge956.gif" alt="mgr" align="MIDDLE" BORDER="0">mol/l of 24/xxlarge946.gif" alt="beta" align="MIDDLE" BORDER="0">-endorphin and maximal release (35% of total) at 20 24/xxlarge956.gif" alt="mgr" align="MIDDLE" BORDER="0">mol/l. The histamine release process was very rapid and terminated within 30 s at 37°C, and in this sense is very similar to the histamine release induced by compound 48/80 or neurotensin. The histamine release was temperature-dependent showing an optimum release around 30°C, and it was independent of available extracellular calcium, but was inhibited in the presence of high extracellular calcium concentrations. Naloxone, only in very high concentrations (10 mmol/l), inhibited the release, and the very same concentration also inhibited the neurotensin — as well as the compound 48/80-induced histamine release. Cromoglycate and benzalkoniumchloride, a 48/80 antagonist, both produced a progressive dose-dependent inhibition of 24/xxlarge946.gif" alt="beta" align="MIDDLE" BORDER="0">-endorphin-, neurotensin- as well as compound 48/80-induced histamine release. Taken together, the findings indicate that the opioid peptide 24/xxlarge946.gif" alt="beta" align="MIDDLE" BORDER="0">-endorphin induces a selective, energy-dependent release of histamine from peritoneal rat mast cells. The pattern of release has much in common with that of compound 48/80 and other basic peptides, such as neurotensin and substance P. In addition this pattern of release is similar to that induced by dynorphin. Send offprint requests to Anita Sydbom at the above address     

Clinical utility of an enhanced human immunodeficiency virus type 1 p24 antigen capture assay

The presence of p24 core antigen in the serum of individuals with human acquired immunodeficiency syndrome has been used as one of the important prognostic markers of HIV-1 infection and also as an end point in evaluating antiviral drugs and vaccines. Unfortunately the majority of p24 antigen present in serum exists as an antigenantibody complex and is not detected with the commercial kits currently available to measure p24 antigen. In this study, we report a simple procedure utilizing treatment of serum samples with glycine buffer (pH 1.85) to dissociate antigen-antibody complexes prior to assaying for p24 antigen. A 300% increase in the number of p24-reactive samples and a 3- to 12-fold increase in the quantity of antigen detected were observed when samples were pretreated with 1.5M glycine buffer (pH 1.85) for 1 hr. Glycine treatment of samples did not result in nonspecific positive tests and samples previously shown to be reactive remained positive. In reconstruction experiments the release of antigen was found to be inversely proportional to the amount of p24 antibody present in the serum. The percentage of HIV-1-infected patients positive for p24 antigen was clearly a function of CD4 count. Forty-nine percent of patients with more than 500 CD4 cells and 100% of patients with less than 200 CD4 were p24 positive. The improved sensitivity for detection of p24 provided by this procedure enhances our understanding of the pathogenesis of AIDS by showing that the majority of patients with HIV-1 infection is p24 positive and facilitates the analysis of data obtained in clinical trials involving anti-HIV compounds.     

HIV核心抗原p24原核系统的高效表达、纯化及活性鉴定

目的　在原核表达系统中对HIVp2 4抗原基因进行克隆和高效表达、纯化并鉴定其活性。方法　利用PCR技术从HIV-1全基因质粒 (B2N)中扩增p2 4抗原基因 ,并克隆入T载体中。通过酶切消化后连接到表达载体pRSET上 ,用此连接产物转化大肠埃希菌BL2 1,经IPTG诱导 ,表达p2 4抗原。利用固定化金属离子 (Ni2 )配体亲和层析技术从表达蛋白中纯化目的蛋白。并运用双酶切技术、SDS-PAGE电泳、WesternBlot(WB)及ELISA法分别对插入基因片段的正确性、表达产物的活性及特异性进行检测。结果　PCR产物约为 6 90bp ,与预期p2 4抗原全基因片段大小一致。重组质粒T-p2 4和pRSET-p2 4经BamHⅠ和HindⅢ双酶切 ,其插入的外源基因片段均为 6 90bp。将纯化前与纯化后的蛋白作SDS PAGE电泳 ,均可见一条约 2 4× 10 3的外源基因表达带 ,与计算的相对分子质量相符。经WB和ELISA试验 ,证明基因工程表达的p2 4抗原具有较高的特异性及活性。结论　成功构建了HIVp2 4表达载体pRSET-p2 4 ,并在原核细胞中高效表达 ,其表达产物具有良好的特异性及活性