新鲜羊膜致I型超敏反应的变应原性研究

研究新鲜人羊膜的变应原性及其致敏后发生I型超敏反应的可能性。建立豚鼠全身主动过敏实验模型。分新鲜羊膜组、新鲜蛋清组(阳性对照)和PBS液组(阴性对照),每组10只豚鼠。观察豚鼠在致敏期和激发后的反应,采用化学荧光法检测外周血组胺含量,血液流变分析系统检测4项血液流变学指标(全血高切变率黏度、全血低切变率黏度、血浆黏度、红细胞聚集指数)。致敏期间各组豚鼠的体重变化无明显差别(P>0.05);激发后羊膜组豚鼠与阴性组表现一致,无异常反应;羊膜组外周血组胺含量及4项血液流变学指标均与阴性对照无明显差别(P>0.05),与阳性对照有显著性差异(P<0.01)。经规范化无菌处理后的新鲜羊膜,一般不具有变应原性,不会引起I型超敏反应。     

柯萨奇B4病毒非结构蛋白P2C重组质粒的构建

目的：构建柯萨奇B4病毒非结构蛋白P2C重组质粒。方法：用RT-PCR技术，从柯萨奇B4病毒中钓取非结构蛋白P2C cDNA片段，克隆入PUCan-T载体，转化大肠杆菌DH5α，构建重组质粒。结果：以柯萨奇B4病毒的总RNA为模板，扩增出987bp大小的片段，克隆入PUCan-T载体，用BamHⅠ、HindⅢ双酶切鉴定，与非结构蛋白P2C的PCR扩增产物片段大小一致。结论：成功地构建了柯萨奇B4病毒非结构蛋白P2C重组质粒。     

Detection of herpesvirus cervicovaginitis by a sequential Papanicolaou--immunoperoxidase technique

Herpes simplex virus (HSV) infection constitutes an immediate threat to the neonate of pregnant women who deliver vaginally, and thus requires a rapid, specific means of diagnosis that is easily applicable to cervicovaginal smears. We applied the peroxidase-antiperoxidase technique to variously fixed, previously stained Papanicolaou smears from 60 women with HSV and 18 negative controls using an HSV-2 antibody and either diaminobenzidine (DAB) or aminoethylcarbazol (AEC) as the chromogen and Mayer's, Gill's, or Lillie-Mayer's hematoxylin as the counterstain. With Papanicolaou stain alone, there was adequate cytologic demonstration of either single cells in aggregates (11%), syncytial giant cells (40%), or both (49%) that displayed a ground-glass appearance (68%), discrete nuclear inclusions (5%), or both (28%). With the peroxidase-antiperoxidase technique, 57 of the 60 HSV specimens (95%) stained moderately or strongly positive for HSV-2, as did sections of herpetic encephalitis and esophagitis. There was no false-positive staining in any of the 18 control smears revealing koilocytosis, Chlamydia, or nonspecific vaginitis. Positivity of the immunostain was more vivid and evenly dispersed through-out the cytoplasm with AEC than with DAB, but tended to wash away with alcohol-based counterstaining. In contrast, DAB was more stable, but was positive predominantly at the cell periphery and cytoplasmic processes. Lillie-Mayer's stain provided the best counterstaining for the cytologic visualization of virocytes and accompanying inflammatory and epithelial cells, which revealed a minimal degree of atypia. The fixative used had no influence on the frequency or degree of immunopositivity of virocytes, but wet fixation with 95% alcohol or Carbowax led to less background staining than Spray-Cyte.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of Behavioral Control and Type A Behavior on Cardiovascular Responses

The purpose of this study was twofold: (a) to examine the relationship between degree of behavioral control and cardiovascular changes in 60 adult males performing an anagram solution task, and (b) to assess the interaction between behavioral control and type of subject (A vs. B). Three conditions were used, differing from each other in the percentage of solvable anagrams (100, 50, and 30%). Systolic blood pressure (SBP), diastolic blood pressure (DBP), and heart rate (HR) were measured before, during, and after the anagram task. In all conditions, during-task SBP, DBP, and HR were significantly elevated above baseline. Blood pressure (BP) elevations appeared to be curvilinearly related to degree of behavioral control, i.e., more elevated in the 50% than in the 100% and 30% conditions. HR changes did not differ among conditions. Concerning the Type A behavior pattern, the results revealed significantly greater BP elevations for Type A than for Type B subjects, especially in the 50% condition, thereby confirming that the physiologic correlates of Type A behavior are predominantly evoked in challenging circumstances. Findings were discussed in the light of results of cardiovascular and endocrinological research. Consideration was given to the possible role of disengagement in the 30% condition and to differences in the pattern of responding between As and Bs.     

Genetic analysis of the formation of the Ysc-Yop translocation pore in macrophages by Yersinia enterocolitica: role of LcrV, YscF and YopN

The Ysc-Yop type III secretion (TTS) system allows extracellular Yersinia bacteria, adhering to eukaryotic target cells, to inject Yop effector proteins in the cytosol of these cells. The secretion apparatus, called the injectisome, ends up with a needle-like structure made of YscF. YopN, one of the proteins secreted by the injectisome is thought to act as a plug. YopB, YopD and LcrV, three other proteins secreted by the injectisome and called 'translocators' form a pore allowing translocation of the Yop effectors across the target cell plasma membrane. Here, we tested the role of LcrV, YscF and YopN in the formation of this pore in macrophages by monitoring the release of the low-molecular-weight fluorescent dye BCECF (2',7'-bis-(2-carboxyethyl)-5-(and-6)-carboxyfluorescein, acetoxymethyl ester, 623Da) and of the high-molecular-weight lactate dehydrogenase (LDH, 135 kDa). BCECF is released through the translocation pore itself provided no Yop effector is trafficking through the channel. In contrast, LDH is released by the osmotic lysis of the target cell that occurs after pore formation. This release is reduced by the GAP activity of YopE. In order to study the role of LcrV, one has to circumvent the regulatory effect of LcrV on the synthesis of YopB and YopD. We observed here that this regulatory role of LcrV is lost in a yopQ mutant and hence we studied the role of LcrV in a yopQ mutant background. A lcrV, yopQ double mutant was deficient in pore formation while able to produce YopB and YopD. Pore formation was restored by the introduction of lcrV(+) but not yopQ(+) confirming that LcrV itself is directly required for pore formation. Bacteria secreting only YopB, YopD and LcrV could form pores, showing that YopB, YopD and LcrV are sufficient for pore formation provided they are secreted by the same bacterium. LcrV is not involved in secretion of YopB and YopD as suggested previously. Bacteria producing normal Ysc injectisomes, including the YscF needle but no translocators did not form pores, indicating that the needle is not sufficient by itself for pore formation, as was also suggested. yopN mutant bacteria formed needles and released BCECF even if they secreted the effectors. This observation suggests that many translocation pores are not filled in the absence of YopN and thus that YopN might form a link between the needle and the pore, guiding the effectors.     

Analysis of children with type 1 diabetes in Korea: high prevalence of specific anti-islet autoantibodies, immunogenetic similarities to Western populations with "unique" haplotypes, and lack of discrimination by aspartic acid at position 57 of DQB

This study analyzed the expression of anti-islet autoantibodies and HLA-DR and -DQ genotypes in Korean children with type 1 diabetes mellitus (T1DM). The positivity of the anti-ICA512, anti-GAD65, and anti-insulin autoantibodies in the newly onset T1DM patients (n = 15) was 66.7%, 86.7%, and 46.7%, respectively, and all of them had one or more of the autoantibodies. HLA analysis showed higher frequencies of HLA-DRB1*0301, *0405, *09012 and -DQB1*0201, *0401, *03032 alleles in T1DM patients compared to controls (P(c) < 0.05). Because HLA-DQB1*0401, *03032 alleles carry aspartic acid at position 57 of DQB, susceptibility to T1DM in Korean children was not related to the presence of aspartic acid at position 57 of DQB1 locus. We suggest this unique HLA-DR, -DQ allele distribution might be an important factor for the low incidence of T1DM in Korea, and the combined anti-islet autoantibody assays could be valuable screening markers for the early detection of T1DM in Korea.     

Mast cell–derived IL-13 downregulates IL-12 production by skin dendritic cells to inhibit the TH1 cell response to cutaneous antigen exposure

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The location and distribution of type A and type B atrial endings in cats

Summary In the attempt to explain the difference in discharge pattern of atrial endings, 131 endings were localized by punctate stimulation, 44 were type A, 77 type B and 10 of an intermediate type. All were located on the dorsal wall of the atria with none on the ventral wall or in the appendage. On the right side, 74% of type A were located in the atria and 63% of type B in or near the veins. On the left side, 67% of type A and 94% of type B were located in or near the veins. Thus, there appeared to be some difference in the location of type A and type B endings on the right side, but on the left side both types of endings were for the most part confined to the venous region. Further, on both right and left sides, these endings were present both in the central part of the atria and in or adjacent to veins. This leads to the suggestion that the difference in discharge patterns is not caused by the location but may be due to some other reasons, e. g. difference of arrangement in the atrial wall with respect to the contractile elements.     

Stimulation by hCRF of C-peptide release in type 2 diabetics during concomitant opioid receptor blockade

Summary Administration of synthetic human corticotropin-releasing factor (hCRF, 2 µg/kg body weight) during simultaneous application of the opioid antagonist naloxone (1.6 mg i.v. bolus, followed by an infusion at a rate of 1.2 mg/h) produced a significant increase in plasma C-peptide levels of six male Type 2 diabetic patients which even exceeded the postprandial values. This stimulatory effect of hCRF/naloxone on plasma C-peptide was less pronounced in six healthy men. hCRF alone did not provoke any reaction of plasma C-peptide in either group.The possibility of a paracrine, CRF-dependent mechanism in pancreatic islets which somehow involves inhibitory opioid receptors is preferentially discussed. Such a mechanism may underlie the stimulatory action of hCRF/naloxone on B cells and would explain the absent reaction of peripheral venous plasma C-peptide to hCRF alone as well as the amplifying effect of simultaneous opioid receptor blockade.Abbreviations ACTH adrenocorticotropic hormone - C-peptide connecting-peptide - CRF corticotropin-releasing factor - hCRF human CRF - oCRF ovine CRF - min minutes - S.D. standard deviation - S.E.M. standard error of the mean This study was supported by the Deutsche Forschungsgemeinschaft (Go 299/3-2)Dedicated to Professor Dr. N. Zöllner on the occasion of his 65th birthday