Structural Analysis of the Complex between Penta-EF-Hand ALG-2 Protein and Sec31A Peptide Reveals a Novel Target Recognition Mechanism of ALG-2

ALG-2, a 22-kDa penta-EF-hand protein, is involved in cell death, signal transduction, membrane trafficking, etc., by interacting with various proteins in mammalian cells in a Ca²⁺-dependent manner. Most known ALG-2-interacting proteins contain proline-rich regions in which either PPYPXnYP (type 1 motif) or PXPGF (type 2 motif) is commonly found. Previous X-ray crystal structural analysis of the complex between ALG-2 and an ALIX peptide revealed that the peptide binds to the two hydrophobic pockets. In the present study, we resolved the crystal structure of the complex between ALG-2 and a peptide of Sec31A (outer shell component of coat complex II, COPII; containing the type 2 motif) and found that the peptide binds to the third hydrophobic pocket (Pocket 3). While amino acid substitution of Phe⁸⁵, a Pocket 3 residue, with Ala abrogated the interaction with Sec31A, it did not affect the interaction with ALIX. On the other hand, amino acid substitution of Tyr¹⁸⁰, a Pocket 1 residue, with Ala caused loss of binding to ALIX, but maintained binding to Sec31A. We conclude that ALG-2 recognizes two types of motifs at different hydrophobic surfaces. Furthermore, based on the results of serial mutational analysis of the ALG-2-binding sites in Sec31A, the type 2 motif was newly defined.     

Noninsulin-dependent diabetes mellitus-related encephalopathy presenting with amnesia, personality change, and autonomic seizure

OBJECTIVE: To determine the knowledge of rubella immune status among practicing obstetrician-gynecologists in the United States and of rubella immunity policies covering healthcare workers in the obstetric-care office setting. DESIGN: Mailed survey questionnaire, August through December 1994. SETTING: Physicians from multiple-practice sites including private office, public institution, university or teaching hospital, and closed panel health maintenance organization settings. PARTICIPANTS: 3,302 practicing obstetrician-gynecologists, chosen by a systematic random sample from the AMA national physician database. MAIN OUTCOME MEASURES: Participants were defined as rubella immune if they reported knowledge of prior rubella vaccination or positive antibody titer. Knowledge of a policy for documenting rubella immunity among employees in the office-based practice setting also was assessed. RESULTS: Questionnaires were returned from 50% (1,666) of the 3,302 surveyed, and 96% (1,599) were evaluable. Approximately 20% (304/1,599) of the responding obstetrician-gynecologists did not have knowledge of documented rubella immunity, and the majority of office-based practices did not require documentation of rubella immunity in the following groups: physicians, 66% (723/1,094); office nurses, 62% (666/1,070); and other office staff, 69% (728/1,063). Sixty-two percent (993/1,599) of responding physicians had individual rubella serologies performed, with 916 known to be positive, 53 reported negative, and 24 reported unknown. Fifty-seven percent (918/1,599) reported receiving monovalent rubella vaccine or trivalent measles-mumps-rubella vaccine. Multiple logistic regression analysis revealed the following to be independent predictors of positive immune status among respondents: female gender (odds ratio [OR], 2.4; 95% confidence interval [CI95], 1.8-3.1), medical school graduation since 1980 (OR, 2.6; CI95, 2.0-3.3), providing obstetric or fertility services (OR, 1.5; CI95, 1.2-1.9), and group practice setting (> or = 5 physicians; OR, 1.2; CI95, 1.1-1.4). CONCLUSIONS: Nationally, nearly one of every five practicing obstetricians may not have documented rubella immunity, and the majority of office-based practices have no system for assuring such immunity. Rubella immunity should extend beyond the hospital setting, with consideration for requiring rubella immunity as a condition for employment. Methods for effective implementation and documentation of current guidelines need to be addressed, particularly in the office setting.     

Circulating vascular cell adhesion molecule-1 (VCAM-1) in atherosclerotic NIDDM patients

Vascular cell adhesion molecule-1 (VCAM-1) has been shown to be highly expressed in atherosclerotic lesions. Although the soluble form of VCAM-1 (sVCAM-1) is detected in human sera, the relation between the degree of atherosclerosis and serum sVCAM-1 level has not been defined. In the present study, sVCAM-1 concentrations were measured in sera from 101 Japanese NIDDM patients. The mean +/- SD serum sVCAM-1 concentration in 26 patients with symptomatic atherosclerotic vascular diseases (789 +/- 187 ng/ml) was higher than that in 75 patients without the disease (664 +/- 175 ng/ml). Among the 101 NIDDM patients, 56 had atherosclerotic change of the carotid arteries, based on the evaluation by high-resolution B-mode ultrasonography. Their sVCAM-1 level was 759 +/- 201 ng/ml, higher than that in 45 patients without any detectable atherosclerosis of the carotid arteries (619 +/- 130 ng/ml). In addition, there was a positive correlation between sVCAM-1 concentration and thickness of the intimal plus medial complex (IMT) of the carotid arteries in the NIDDM patients (r = 0.41, P < 0.0001). Multivariate regression analysis revealed significant predictors of mean IMT value to be sVCAM-1 concentration (F = 62.88, P = 0.0001) and age (F = 9.59, P = 0.0026). By contrast, sVCAM-1 concentration was not increased in nondiabetic patients with atherosclerotic change of the carotid arteries (668 +/- 191 ng/ml; n = 36) compared with those without the atherosclerotic change (632 +/- 177 ng/ml; n = 28), and there was no correlation between sVCAM-1 level and IMT of the carotid arteries in the nondiabetic subjects. These results indicate that circulating sVCAM-1 may be a marker of atherosclerotic lesions in NIDDM patients with symptomatic and asymptomatic atherosclerosis.     

An ecological perspective on smoking among Asian American college students: The roles of social smoking and smoking motives.

Using electronic diaries, the present study examined the roles of social smoking and smoking motives in relation to cigarette use patterns among Asian American college smokers. Multilevel modeling results showed that participants smoked more cigarettes when smoking with peers than when smoking alone. Participants' coping (but not social) motives moderated the within-person associations between smoking with peers and the cigarettes smoked during a smoking episode. The findings support the utility of an ecological perspective in examining the dynamic interaction between smoking motives and the social settings of cigarette use, and call for further research on the social smoking behaviors in diverse populations. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Macrophage Activation Markers Predict Liver-Related Complications in Primary Biliary Cholangitis

Primary biliary cholangitis (PBC) has a wide variation in clinical presentation and course. There is no significant correlation between these symptoms and the disease stage, although patients with more advanced stages generally have more symptoms. It is important to develop biomarkers in order to identify patients with an increased risk of complications and end-stage liver disease. This study investigated surrogate markers for risk estimation of PBC-related complications, including a study population of 77 patients with PBC who underwent liver biopsy and were measured for serum levels of macrophage activation markers, soluble CD163 (sCD163), soluble mannose receptor (sMR), and zonulin. Patients with PBC were divided into symptomatic (Group S, n = 20) and asymptomatic (Group A, n = 57) groups. The correlations of histological stages based on both Scheuer and Nakanuma classifications with the three serum markers were investigated. The Nakanuma classification involves grading for liver fibrosis and bile duct loss. The three biomarkers were assessed for their diagnostic ability to identify patients with PBC having high risk of developing complications. The predictive factors of these complications were examined as well. Group S had significantly higher serum sMR (p = 0.011) and sCD163 (p = 0.048) levels versus Group A. A composite index of sMR and sCD163 measurements had significantly better prediction performance than sCD163 alone (p = 0.012), although not when compared to sMR alone (p = 0.129). Serum sMR was an independent factor for developing complications on both univariate (Odds ratio (OR) = 30.20, 95% confidence interval (95% CI): 3.410–267.0, p = 0.00220), and multivariate (OR = 33.70, 95% CI: 3.6600–311.0, p = 0.0019) analyses. Patients with PBC having sMR of ≥56.6 had a higher incidence of clinical complications versus those with a sMR of <56.6. Serum sMR predicts the development of complications in patients with PBC. sMR plus sCD163 showed better predictive power than either marker alone, although the addition of sCD163 did not improve the predictive power of sMR. Future prospective studies are required in order to validate the findings of the present study.     

Comparison between Er:YAG laser and conventional technique for root caries treatment in vitro

Effective ablation of dental hard tissues by means of the erbium-doped:yttrium-aluminum garnet (Er:YAG) laser has been reported recently, and its application to caries removal and cavity preparation has been expected. However, few studies have investigated the capability of the Er:YAG laser to treat caries. In the present study, the effectiveness of caries removal by using an Er:YAG laser in vitro was compared with that of conventional mechanical treatment. Thirty-one extracted human teeth with root caries were used. Half of the caries in each tooth was treated with the Er:YAG laser, and the other was removed with a conventional bur or was left untreated as a control. Laser treatment was performed by means of a combination of contact and non-contact irradiation modes with cooling water spray, with a new fiber delivery and contact probe system. Conventional bur treatment was conducted by means of a low-speed micromotor. Measurements of the time required for caries removal, histopathological observations of decalcified serial sections, scanning electron microscope (SEM) observations, and hardness measurements of the treated cavity-floor dentin were performed for each treatment. Due to the careful irradiation technique, a longer treatment time was required for the complete removal of carious dentin by the Er:YAG laser. However, the Er:YAG laser ablated carious dentin effectively with minimal thermal damage to the surrounding intact dentin, and removed infected and softened carious dentin to the same degree as the bur treatment. In addition, a lower degree of vibration was noted with the Er:YAG laser treatment. The SEM examination revealed characteristic micro-irregularities of the lased dentin surface. Our results show that the Er:YAG laser system is promising as a new technical modality for caries treatment.     

Evidence supporting the activity of 2'-C-cyano-2'-deoxy-1-beta-D-arabino-pentafuranosylcytosine as a terminator in enzymatic DNA-chain elongation

To investigate the stability of 2'-C-cyano-2'-deoxy-1-beta-D-arabino-pentafuranosylcytosine 3'-phosphoric acid, its thymidine ester was prepared via the phosphoramidite method using allyl protection for the phosphate function. This ester is stable under acidic conditions but extremely labile under basic conditions, decomposing with a cleavage of the internucleotide bond even at pH 7.3.