Differential scanning calorimetric study of the complexes of modified myosin subfragment 1 with ADP and vanadate or beryllium fluoride

Summary The effects of various modifications of rabbit skeletal myosin subfragment 1 on thermal denaturation of subfragment 1 in ternary complexes with Mg-ADP and orthovanadate (V_i) or beryllium fluoride (BeF_x) have been studied by differential scanning calorimetry. It has been shown that specific modifications of SH₁ group of Cys-707 by different sulfhydryl reagents, trinitrophenylation of Lys-83, and reductive methylation of lysine residues promote the decomposition of the S1·ADP·V_i complex and change the character of structural transitions of the subfragment 1 molecule induced by the formation of this complex, but they have much less or no influence on subfragment 1 thermal stability in the S1·ADP·BeF_x complex. Thus, the differential scanning calorimetric studies on modified subfragment 1 preparations reveal a significant difference between S1·ADP·V_i and S1·ADP·BeF_x complexes. It is suggested that S1·ADP·V_i and S1·ADP·BeF_x complexes represent structural analogues of different transition states of the ATPase cycle, namely the intermediate states S1^**·ADP·P_i and S1^*·ATP, respectively. It is also proposed that during formation of the S1·ADP·V_i complex the region containing both Cys-707 and Lys-83 plays an important role in the spread of conformational changes from the active site of subfragment 1 ATPase throughout the structure of the entire subfragment 1 molecule. In such a case, the effects of reductive methylation of lysine residues on the subfragment 1 structure in the S1·ADP·V_i complex are related to the modification of Lys-83.     

Demonstration of immunoglobulin on the surface of thymus lymphocytes

Immunoglobulin molecules on the surface of mouse thymus cells have been shown by immunofluorescence. Ninety-five to 100 per cent of thymus lymphocytes were found to bind polyvalent rabbit anti-mouse immunoglobulin, although the density of the fluorescence was much less than that on the surface of B lymphocytes derived from the spleen.

Two purified antisera, an anti-IgM and an anti-kappa chain serum, also stained the cells.

The resynthesis of these immunoglobulin molecules in vitro was demonstrated after they had been removed with pronase.

     

Rosiglitazone, an agonist of peroxisome proliferator-activated receptor γ, reduces pulmonary inflammatory response in a rat model of endotoxemia

Objective: The effect of rosiglitazone, a potent peroxisome proliferator-activated receptor γ (PPAR-γ) agonist, on pulmonary inflammation in endotoxemia was investigated. Materials and methods: Male Wistar rats were given either lipopolysaccharide (LPS, 6 mg/kg i.v.) or saline, pretreated with rosiglitazone (0.3 mg/kg i.v.) or its vehicle (dimethyl sulphoxide) 30 min before LPS. The selective PPAR-γ antagonist GW9662 (0.3 mg/kg i.v.) was given 20 min before rosiglitazone. Wet/dry weight (W/D) ratio, myeloperoxidase (MPO) activity, malondialdehyde (MDA) as well as TNF-α and CINC-1 concentrations were measured in lung tissues 4 h after LPS injection. Expression of ICAM-1, NF-κB p65 and PPAR-γ were also determined by immunohistochemistry or Western blot analysis. Results: Rosiglitazone pretreatment significantly attenuated the increases in W/D ratio, MPO activity and MDA levels, and reduced pulmonary overproduction of TNF-α and CINC-1 as well as expression of ICAM-1 following endotoxemia. Rosiglitazone also inhibited the nuclear localization of NF-κB and up-regulated the expression of PPAR-γ protein. The specific PPAR-γ antagonist GW9662 abolished the effect of rosiglitazone. Conclusion: These findings suggest that PPAR-γ agonists might be used as therapeutic agents in the therapy of inflammatory lung injury related to endotoxemia. Received 8 January 2005; returned for revision 6 July 2005; returned for final revision 20 July 2005; accepted by M. Katori 31 July 2005     

神经导航中脑组织变形补偿的点云处理

有限元方法是解决神经导航中脑组织变形的重要方法,需要手术过程中的脑皮层信息作为其边界条件.本文通过非结构点云进行了脑皮层信息的表示,并通过对其进行处理来获取有限元方法的边界条件.点云处理包括纹理映射、分割、简化和去噪,其中非结构点云的简化与去噪采用了改进的基于表面特性k邻域的聚类方法.实验结果证明所采用的点云处理方法是可靠的.     

Expression and characterization of a soluble rubella virus E1 envelope protein

Individual specific antigenic rubella virus (RV) structural proteins are required for accurate serological diagnosis of acute and congenital rubella infections as well as rubella immune status. The RV envelope glycoprotein E1 is the major target antigen and plays an important role in viral-specific immune responses. The native virion is difficult to produce in large quantities and the protein subunits are also difficult to isolate without loss of antigenicity. The production of a soluble RV E1 (designated E1ΔTm) using the baculovirus-insect cell expression system is described. In contrast to wild-type RV E1, the genetically engineered E1ΔTm protein lacks a transmembrane anchor. It behaved as a secretory protein and was secreted abundantly from insect cells. Pulse-chase studies were used to examine the synthesis, glycosylation, and secretion of E1ΔTm by the insect cells. The secreted E1ΔTm protein was purified from serum-free medium by onestep immunochromatography. The purified E1ΔTm protein retained full antigenicity and may be a convenient source of E1 protein for use in diagnostic assay and rubella vaccine development.     

Working despite pain: Factors associated with work attendance versus dysfunction

A cross-sectional investigation of psychosocial variables in 63 female employees matched for experienced pain was conducted to study the difference between back pain sufferers who were working (Copers) and those who were off work (Dysfunctional). The subjects reported moderate to severe pain often or always during the past year and were employed at the same hospital. Thirty-seven women who had not been off work for pain made up the Copers group, whereas 26 women who had been off work for their pain made up the Dysfunctional group. Subjects were interviewed and completed a battery of questionnaires designed to penetrate level of dysfunction, perceived health, work and social satisfaction, perceived workload, coping strategies, and pain beliefs. Multiple covariate analyses that controlled for perceived workload, smoking, low-back mobility, and obesity revealed significant differences between the groups on levels of functioning, pain beliefs, and coping strategies used. Dysfunctional subjects had stronger beliefs that pain was directly related to activities that they had little control over their pain, that their health was poor, and that they tended to focus more on their pain. A discriminant analysis correctly classified 83% of the subjects as to work status based on six psychosocial variables. These results not only demonstrate the importance of psychosocial factors in back pain, but underscore the fact that work absence for back pain may he controlled by psychological factors related to beliefs and coping strategies. Future research may attempt to use these factors in the screening of patients.     

Evaluation of dry and wet transported intravaginal swabs in detection of Chlamydia trachomatis and Neisseria gonorrhoeae infections in female soldiers by PCR

Screening women for sexually transmitted diseases (STD) in nonclinic settings is highly desirable because many infections are asymptomatic. This is especially true for military women, for whom logistical, social, and other job-related obstacles present barriers to accessing medical care. We assessed the accuracy of intravaginal swabs transported by mail in a wet versus a dry state for PCR (Amplicor CT/NG test) detection of chlamydia and gonorrhea infections in a cross-sectional study of 793 active-duty military women attending an STD clinic. PCR tests of vaginal swabs (wet and dry) were compared to local clinical methods used on cervical swabs. Standard wet vaginal swab PCR testing detected more chlamydia (11.6%) than cervical enzyme immunoassay (9.3%). For detection of chlamydia using wet swabs, the sensitivity and specificity compared with adjudicated true positives were 94.6% (87 of 92) and 99.3% (696 of 701), respectively. Comparing dry swabs to true-positives for chlamydia, the sensitivity was 91.3% (84 of 92) and the specificity was 99.3% (696 of 701). Standard wet vaginal swab PCR detected more gonorrhea (3.3%) than routine cervical culture (2.1%). The sensitivity and specificity of PCR testing of wet swabs compared to true-positives (infected patients) were 96.3% (26 of 27) and 98.2% (752 of 766) for gonorrhea, respectively. For gonorrhea, the sensitivity and specificity of dry swabs compared to true-positives (infected patients) were 88.9% (24 of 27) and 98.3% (753 of 766), respectively. PCR testing of wet and dry transported intravaginal swabs to detect chlamydia and gonorrhea infections was an accurate diagnostic method for military women.