Identification of wild-type and mutant p53 peptides binding to HLA-A2 assessed by a peptide loading-deficient cell line assay and a novel major histocompatibility complex class I peptide binding assay

Mutations of the p53 gene are the most frequently observed genetic changes in human cancers; often leading to an overexpression of the wild-type (wt) p53 protein. Demonstrable T cell reactivity against tumor cells overexpressing wt or mutant p53-derived peptides could support the application of such epitopes in cancer immunotherapies. As the binding of peptide to MHC class I molecules is a prerequisite for antigen-specific T cell recognition, we evaluated the ability of wt and mutant p53 peptides to bind to HLA-A2.1 using two independent flow cytometry-based assay systems, the T2 major histocompatibility complex (MHC) class I peptide stabilization assay (stabilization assay) and the peptide-induced MHC class I reconstitution assay (reconstitution assay). The twenty selected wt sequences each conformed to the previously reported HLA-A2.1 peptide binding motif. Seven of the wt p53 and 2/13 mutant p53 peptides derived from the previously chosen wt peptides bound to HLA-A2.1 in both the stabilization and the reconstitution assays. An additional six wt and six mutant p53 peptides, presumably exhibiting lower affinity for HLA-A2.1, were identified only in the reconstitution assay. Those p53 peptides binding HLA-A2.1 may provide useful immunogens for the generation of HLA-A2.1-restricted cytolytic T lymphocytes in vitro and in vivo.     

REACTIVITY OF LYMPHOCYTE SUBPOPULATIONS IN HUMAN MIXED LYMPHOCYTE CULTURE

Loss of antigenicity in the mixed lymphocyte culture (MLC) reaction of lymphocytes precultured at 22°C for 7–10 days was accompanied by a decrease in bone-marrow-derived lymphocytes (B cells) from 22 ± 1% to 13 ± 1%, and an increase in thymus-derived lymphocytes (T cells) from 65 ± 2% to 83 ± 1% (P < 0.001). Depletion of B cells from a fresh lymphocyte suspension by either antihuman immunoglobulin-coated column fractionation or by sheep red blood cell (SRBC) rosette formation resulted in a significant reduction of the cell's ability to stimulate in MLC (P < 0.001). Coating of lymphocytes with rabbit antihuman brain serum abrogated their ability to respond but not the ability to stimulate in MLC.     

Poly(2,5-dialkoxy-p-phenylene)s—synthesis and properties

The synthesis of poly(2,5-dialkoxy-p-phenylene)s (PPP) by palladium-catalyzed polycondensation of 2,5-dialkoxy-4-bromophenylboronic acids is described. Number-average degrees of polymerization of ca. 30 have been reached at best. The characterization of the polymers by NMR, IR, UV-VIS and fluorescence spectra, data on the bulk structure obtained by X-ray methods, melting behavior and thermal stability are reported. PPP's with long alkoxy side chains exhibit a layered phase structure, those with short or branched alkoxy side chains, cylindrical packing. Poly(2,5-dibutoxy-p-phenylene) was prepared with chain lengths 7 ≤ P_n ≤ 30. The polymer with P_n of 30 shows thermal transitions in the bulk phase at 182°C and 227°C. It forms an anisotropic melt above 227°C which only starts decomposing if heated above 300°C. The polymers of shorter chain length show isotropic melts, with the melting point strongly depending on chain length.     

Determination of Dissolved Oxygen in Heterogenous Systems Particularly in Emulsions and Oily Liquids

The content of dissolved oxygen was determined by four independent methods in a series of non-aqueous or heterogenous systems. The Lex-O₂-Content Analyzer represents a fast and simple apparatus that employs a coulometric oxygen assay with Hersch cell detection. A comparison of the results with different methods demonstrates the reliability of the Lex-O₂ in the determination of oxygen dissolved in heterogeneous or non-aqueous systems. Therefore, this apparatus can be recommended for the measurement of oxygen in oxygenator or perfusion fluids, as well as in blood substitutes or other oxygen transporting systems.     

Early biochemical and morphological changes of the rat adrenal medulla induced by xylitol

Long-term administration of high doses of xylitol and other polyols in rats has been associated with an increase in adrenal medullary hyperplasia and neoplasia. In order to exclude age-related factors and to differentiate between unspecific stress reactions and direct effects of the compound administered, a model was developed for quantifying early adrenomedullary responses. Male SD rats were fed xylitol (10% or 20% in the diet) for 2 and 8 weeks, and early biochemical changes were correlated with a stereological analysis of the adrenal medulla. At first, the in vivo rate of catecholamine (CA) biosynthesis was slightly decreased (at 2 weeks). This was followed by an increase in dopamine--hydroxylase (DBH) activity (at 8 weeks). By that time, the total chromaffin cell volume had increased and the number of chromaffin cells per reference volume had decreased in a dose-dependent way. The total number of chromaffin cells per adrenal gland showed a distinct tendency towards an increase. Adrenal epinephrine and norepinephrine contents were not altered, and both tyrosine hydroxylase and phenylethanolamine-N-methyltransferase activities remained unchanged. These data suggest that continued xylitol administration evoked an inhibitory effect on CA synthesis that, together with stimulation of the adrenal medulla brought about by the compound, resulted in compensatory medullary hypertrophy and hyperplasia.     

Significance of laboratory diagnostic study parameters of the hepatobiliary system in the evaluation of cardiovascular diseases--results of cross sectional and longitudinal studies in patients with suspicious cardiovascular findings in an epidemiologic follow-up study

With the help of an approx. 280 000 inhabitants check of x-ray-morphological suspects of a heart disease the laboratory parameters thymol turbidity test, zinc-sulphate turbidity test, total protein, serum protein electrophoresis and the enzymes "ASAT" and "ALAT" in longitudinal section and cross-section were examined in an intervention study during 5 years. All these parameters show standard values on the average, but compared to a healthy reference they showed significant differences, which relations to heart and vessel diseases could be deduced from. This referred especially to age-inverse behaviour of the transaminases only concerning male test persons. The examination of the longitudinal section confirmed exactly the results of the cross-section analysis. On the one hand it underlines the results got from the cross-section and in the second place it contributes to the efficient diagnostics by learning undone a repetition of these laboratory examinations during the period of 5 years with such a defined population. The relations of the particularities mentioned obviously refer to subclinical fields. Effects of therapy do not reflect in the laboratory findings.     

Investigations on neurotoxicity of chemical substances at the workplace

Summary A cross-sectional study was performed in order to investigate the influence of chronic lead-exposure on the peripheral nervous system. We examined 148 male workers of a storage battery manufacturing plant, who had been exposed to lead metal and inorganic lead compounds for 1 to 28 years (mean 11 years). Fifteen workers with non-occupational risks of peripheral neuropathy (former diseases, alcohol abuse, medication) were excluded from the study. The investigation program comprised: case history, physical examination, analyses of blood- and urine-samples and determination of maximal motor, mixed and sensory conduction velocity (NCV) of the ulnar and median nerve of the right forearm. Objectively no worker showed any signs of health effects related to lead exposure. The Biological Monitoring included the determination of (1) Blood-lead level (Pb-B), (2) Free erythrocyte porphyrins (FEP), (3) -Aminolevulinic acid dehydratase (ALA-D) and (4) -Aminolevulinic acid in urine (ALA-U). Further time-weighted-average (TWA)-values of Pb-B were calculated on the basis of several determinations over the period 1975–1981. The following actual (TWA) median values resulted: Pb-B 53 g/dl (54 g/dl), ALA-U 5.6 mg/l (8.4 mg/l), FEP 2.0 mg/l (2.0 mg/l). The Biologischer Arbeitsstoff Toleranz Wert (BAT) of 70 g//dl for Pb-B was exceeded in 15 workers (11%), and of 15 mg/l for ALA-U in 30 cases (23%). In comparison with age-matched controls, the lead workers showed a mild slowing of NCV with mean values between 0.8 and 2.0 m/s. Multiple stepwise regression analyses revealed statistically significant correlations between the four NCV and age as well as Pb-B. There were better correlations by using TWA than actual data of Pb-B. Consideration of the results of the regression analyses, together with an evaluation of the individual neurophysiological status as a function of internal lead exposure, a dose-effect-relationship was found only in the case of Pb-B exceeding 70 g/dl. From our study it is concluded that chronic lead exposure resulting in blood-lead levels of below 70 g/dl is no occupational risk causing a functionally significant slowing of nerve conduction velocities.With Grants from the Deutsche Forschungsgemeinschaft, Bonn (Project no. Va 23/19-1)     

胰腺切除术后出血的诊断与治疗：669例连续胰腺切除术分析

目的：分析胰腺切除术后出血（postpancreatectomy hemorrhage，PPH）的临床病程及预后。背景：PPH是最致命的胰腺术后并发症，但目前仍缺少其标准治疗规程。方法：1992年至2006年，对1524例施行胰腺手术的患者进行了前瞻性研究。根据以下指标进行PPH危险分级：PPH严重程度（轻度，即血红蛋白浓度下降〈3g／dL；重度，即血红蛋白浓度下降〉3g／dL），PPH出现时间（早期，即术后1～5天；晚期，术后第6天），合并胰瘘，肠道内或肠道外出血表现以及存在“复杂的”血管病理变化（腐蚀、假性动脉瘤）。     

Interleukin-6 and selected plasma proteins in healthy persons of different ages

In the present study, interleukin-6(IL-6) and several acute phase proteins were measured in healthy participants (23–87 years of age). A linear correlation between IL-6 and age was established with an increase of 0.016 pg/ml(00.004) per year of life. Whereas CRP remained below 0.5 mg/dl in all participants, an increase with age for fibrinogen and an inverse relation for albumin as well as transferrin were obtained. However, the increase of IL-6 did not correlate with any of these changes. IL-6 associated diseases may therefore occur more often with advancing age, but in healthy participants IL-6 does not explain the changing plasma protein pattern resembling that of an acute phase reaction.